Preparation and biological evaluation of viridicatumtoxin analogs

ABSTRACT

In one aspect, the present invention provides novel derivatives of viridicatumtoxin of the formula wherein the variables are as defined herein. The application also provides compositions, methods of treatment, and methods of synthesis thereof.

This application claims the benefit of priority to U.S. ProvisionalApplication Ser. No. 62/027,500, filed on Jul. 22, 2014, the entirecontents of which are hereby incorporated by reference.

This invention was made with government support under Grant NumberAI055475 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This disclosure relates to the fields of medicine, pharmacology,chemistry, antimicrobial activity, and oncology. In particular, newcompounds, compositions, methods of treatment, and methods of synthesisrelating to viridicatumtoxin and derivatives thereof are disclosed.

2. Related Art

Since the discovery of chlortetracycline (4; FIG. 1A) in the late1940's, tetracycline antibiotics such as chlortetracycline (4),oxytetracycline (5), and tetracycline (6) have been commonly prescribedto treat bacterial infections (Duggar, 1948). Throughout the years, asbacterial resistance grew or improved therapeutic properties wereneeded, additional therapeutic agents including second generationtetracycline derivatives such as minocycline (7) and doxycycline (8) andthird generation tetracycline derivatives such as tigecycline (9) anderavacycline (TP-434, 10) have been developed (FIG. 1A) (Tally, et al,1995; Sutcliffe, et al., 2013; Chopra and Roberts, 2001).

Penicillium is a genus of ascomycetous fungi of major importance in thenatural environment as well as food and drug production. Members of thegenus may be best known for producing penicillin, a molecule that isused as an antibiotic, which kills or stops the growth of certain kindsof bacteria inside the body. According to the Dictionary of the Fungi(10th edition, 2008), the widespread genus contains over 300 species.Additionally, the majority of the tetracycline antibiotics havebacterial origins but some tetracycline antibiotics have fungal origins.Tetracycline antibiotics which have fungal origins includeviridicatumtoxin B (1), viridicatumtoxin A (2), spirohexaline (3),hypomycetin (11), anthrotainin (TAN-1652, 12), TAN-1612 (13), andBMS-192548 (14) (FIG. 1B & FIG. 2) (Zheng, et al., 2008; Hutchinson, etal., 1973; Inokoshi, et al., 2013; Breinholt, et al., 1997; Wong, etal., 1993; JP 06-40995; Kodukula, et al., 1995; and Shu, et al., 1995).Several tetracycline antibiotics with fungal origins includingviridicatumtoxin B (1), viridicatumtoxin A (2), and spirohexaline (3)are also structurally unique from the earlier tetracycline derivativesin the structure also contains a spirobicyclic system (ring system EF)derived from a geranyl subunit (Zheng, et al., 2008; Hutchinson, et al.,1973; Inokoshi, et al., 2013) (FIG. 2).

Without being bound by theory, the proposed biosynthetic pathway toproduce the viridicatumtoxin A (2) has been reported and is shown inFIG. 3 (De Jesus, et al., 1982; Chooi, et al., 2010; Chooi, et al.,2012; Chooi, et al., 2013). Due to the biological activity of thesecompounds and the need for multi-gram quantities of the compounds, acommercially scalable synthesis is needed. Since the discovery of thecommercial importance of these molecules for the biological activity,many efforts have been undertaken to synthesize tetracycline derivativesincluding recent efforts by the Myers (Charest, et al., 2005; Charest,et al., 2005; Brubaker and Myers, 2007; Sun, et al., 2008; Kummer, etal., 2011; Wright and Myers, 2011) and Evans groups (Wzorek, et al.,2012).

In 2008, Kim, et al., isolated viridicatumtoxin B (1) from Penicilliumsp. FR11 along with viridicatumtoxin A (2). This compound wasinvestigated through NMR spectroscopy and assigned the structure 1′.These compounds have been shown to have potent antibacterial propertiesin a number of bacterial strains including both gram positive and gramnegative bacteria (Kim, et al., 2008). Without being bound by theory,further study and analysis suggests that the viridicatumtoxin'santibacterial properties arise not by binding to the 30S subunit of theribosome like many tetracycline compounds (e.g., 4-10, FIG. 1A) but byinhibiting UPP synthase, an enzyme associated with bacterialpeptidoglycan biosynthesis (Inokoshi, et al., 2013; Koyama, et al.,2013). Furthermore, viridicatumtoxin A (2) shows promising anticanceractivity against a selection of cancer cell lines (NIH Results ofViridicatumtoxin A NCI 60 Cell line assay) as well as shows antiviralactivity (WO 2009/008906).

As such, new analogs of viridicatumtoxin could provide access to a moreefficacious antimicrobial or cancer drug and new methods of synthesiscould allow cost effective clinical access to these compound for use inthe treatment of microbial infections and as chemotherapeutic agents.

SUMMARY OF THE INVENTION

Thus, in accordance with the present invention, there is provided acompound of the formula:

wherein: X₁ is absent such that atoms 16 and 17 are only connected bythe shown single bond, a covalent bond such that a double bond is formedbetween atoms 16 and 17, —O—, alkanediyl_((C≦8)), or substitutedalkanediyl_((C≦8)); Y₁, Y₂, and Y₃ are each independently alkyl_((C≦12))or substituted alkyl_((C≦12)); R₁ is hydrogen, hydroxy, or oxo, providedthat when R₁ is oxo, the bond between R₁ and atom number 5 is a doublebond and when the bond between R₁ and atom number 5 is a double bondthen R₁ is oxo, alkoxy_((C≦8)), or substituted alkoxy_((C≦8)); R₂ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₃, R₆, R₇, and R₁₀ are each independently selected from: hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; R₄ and R₅are each independently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: X₁ is absent such that atoms 16 and 17 are only connected bythe shown single bond, a covalent bond such that a double bond is formedbetween atoms 16 and 17, —O—, alkanediyl_((C≦8)), or substitutedalkanediyl_((C≦8)); Y₁, Y₂, and Y₃ are each independently alkyl_((C≦12))or substituted alkyl_((C≦12)); R₁ is hydrogen, hydroxy, or oxo, providedthat when R₁ is oxo, the bond between R₁ and atom number 5 is a doublebond and when the bond between R₁ and atom number 5 is a double bondthen R₁ is oxo, alkoxy_((C≦8)), or substituted alkoxy_((C≦8)); R₂ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ and R₅ are each independently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: X₁ is a covalent bond such that a double bond is formed betweenatoms 16 and 17, —O—, alkanediyl_((C≦8)), or substitutedalkanediyl_((C≦8)); R₁ is hydrogen, hydroxy, or oxo, provided that whenR₁ is oxo, the bond between R₁ and atom number 5 is a double bond andwhen the bond between R₁ and atom number 5 is a double bond then R₁ isoxo, alkoxy_((C≦8)), or substituted alkoxy_((C≦8)); R₂ is hydrogen,amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)),alkoxy_((C≦12)), alkylamino_((C≦12)), dialkylamino_((C≦18)), or asubstituted version of any of these groups; R₄ and R₅ are eachindependently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: R₁ is hydroxy, or oxo, provided that when R₁ is oxo, the bondbetween R₁ and atom number 5 is a double bond and when the bond betweenR₁ and atom number 5 is a double bond then R₁ is oxo, alkoxy_((C≦8)), orsubstituted alkoxy_((C≦8)); R₂ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), or a substituted version ofany of these groups; R₄ and R₅ are each independently selected from:hydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein: the linker is alkanediyl_((C≦12)),alkenediyl_((C≦12)), arenediyl_((C≦12)), heteroarenediyl_((C≦12)),heterocycloalkanediyl_((C≦12)) or a substituted version of any of thesegroups; and the biomolecule is a protein, a polypeptide, an antibody, animaging agent, or a small molecule therapeutic agent; R₉ is hydrogen,amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)),alkoxy_((C≦12)), alkylamino_((C≦12)), dialkylamino_((C≦18)),amido_((C≦12)), or a substituted version of any of these groups; or—NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen, alkyl_((C≦8)), orsubstituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8)) or substitutedalkanediyl_((C≦8)); and R₁₄ and R₁₅ are each independently selectedfrom: hydrogen, alkyl_((C≦12)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦12)), or a substituted version of any of these groups; or R₁₄and R₁₅ are taken together and are alkanediyl_((C≦8)),alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or a substituted version ofany of these groups; or a pharmaceutically acceptable salt or tautomerthereof. In some embodiments, the compound is further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), substitutedalkyl_((C≦8)), or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12)) orsubstituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein: the linker is alkanediyl_((C≦12)),alkenediyl_((C≦12)), arenediyl_((C≦12)), heteroarenediyl_((C≦12)),heterocycloalkanediyl_((C≦12)) or a substituted version of any of thesegroups; and the biomolecule is an antibody, an imaging agent, a protein,or a small molecule therapeutic agent; R₉ is hydrogen, amino, carboxy,cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), amido_((C≦12)), or asubstituted version of any of these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅;wherein: R₁₂ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8));R₁₃ is alkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ andR₁₅ are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen or -linker-biomolecule; wherein: thelinker is alkanediyl_((C≦12)), alkenediyl_((C≦12)), arenediyl_((C≦12)),heteroarenediyl_((C≦12)), or heterocycloalkanediyl_((C≦12)); and thebiomolecule is an antibody; and R₉ is hydrogen, amino, carboxy, cyano,halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), amido_((C≦12)), or asubstituted version of any of these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅;wherein: R₁₂ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8));R₁₃ is alkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ andR₁₅ are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, halo, or hydroxy; alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule; and R₉ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), amido_((C≦12)), or asubstituted version of any of these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅;wherein: R₁₂ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8));R₁₃ is alkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ andR₁₅ are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundfurther defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, halo, or hydroxy; alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule; and R₉ is hydrogen, amino, cyano, halo, hydroxy;alkoxy_((C≦12)), amido_((C≦12)), or a substituted version of either ofthese groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof. In some embodiments, the compoundis further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, halo, or hydroxy; alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule; and R₉ is —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ ishydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ isalkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or a pharmaceutically acceptable salt ortautomer thereof. In some embodiments, R₁ is alkoxy_((C≦8)) orsubstituted alkoxy_((C≦8)). In some embodiments, R₁ is alkoxy_((C≦6)).In some embodiments, R₁ is methoxy. In other embodiments, R₁ is oxo. Insome embodiments, R₂ is hydrogen. In other embodiments, R₂ is halo. Insome embodiments, R₂ is fluoro, chloro, bromo, or iodo. In otherembodiments, R₂ is hydroxy. In other embodiments, R₂ is amino. In otherembodiments, R₂ is dialkylamino_((C≦12)) or substituteddialkylamino_((C≦12)). In some embodiments, R₂ is dialkylamino_((C≦12)).In some embodiments, R₂ is dimethylamino or (2-aminoethyl)methylamino.In some embodiments, R₄ is hydrogen. In some embodiments, R₅ ishydrogen. In other embodiments, R₅ isalkanediyl_((C≦8))-heterocycloalkyl_((C≦8)). In some embodiments, R₅ is—CH₂CH₂N(CH₂)₄. In other embodiments, R₅ isalkanediyl_((C≦8))-alkylamino_((C≦8)). In some embodiments, R₅ is—(CH₂)₂NH(CH₂)₄CH(NH₂)CO₂H. In some embodiments, R₈ is hydrogen. Inother embodiments, R₈ is —X₂—R₁₁. In some embodiments, X₂ isalkanediyl_((C≦6)) or substituted alkanediyl_((C≦6)). In someembodiments, X₂ is alkanediyl_((C≦6)). In some embodiments, X₂ is—CH₂CH₂CH₂— or —CH₂CH₂—. In some embodiments, R₁₁ is amino. In otherembodiments, R₁₁ is heterocycloalkyl_((C≦12)), or substitutedheterocycloalkyl_((C≦12)). In some embodiments, R₁₁ is4-N-methyl-piperazinyl. In other embodiments, R₈ is a-linker-biomolecule. In some embodiments, the linker isalkanediyl_((C≦12)), alkenediyl_((C≦12)), arenediyl_((C≦12)),heteroarenediyl_((C≦12)), heterocycloalkanediyl_((C≦12)) or asubstituted version of any of these groups. In some embodiments, thebiomolecule is an antibody, a protein, or a small molecule therapeuticagent. In some embodiments, the biomolecule is an antibody. In someembodiments, R₉ is hydrogen. In other embodiments, R₉ is halo. In someembodiments, R₉ is fluoro or iodo. In other embodiments, R₉ isamido_((C≦12)) or substituted amido_((C≦12)). In some embodiments, R₉ is—NHC(O)CH₂NH₂. In other embodiments, R₉ is —NR₁₂C(O)R₁₃—NR₁₄R₁₅. In someembodiments, R₁₂ is hydrogen. In other embodiments, R₁₂ is alkyl_((C≦6))or substituted alkyl_((C≦6)). In some embodiments, R₁₃ isalkanediyl_((C≦8)). In some embodiments, R₁₃ is —CH₂—. In someembodiments, R₁₄ is alkyl_((C≦12)) or substituted alkyl_((C≦12)). Insome embodiments, R₁₄ is t-butyl. In some embodiments, R₁₅ is hydrogen.In some embodiments, the compound is further defined as:

or a pharmaceutically acceptable salt, tautomer, or optical isomerthereof.

In another aspect, the present disclosure provides a pharmaceuticalcomposition comprising a compound as described herein and an excipient.In some embodiments, the composition is formulated for administration:orally, intraadiposally, intraarterially, intraarticularly,intracranially, intradermally, intralesionally, intramuscularly,intranasally, intraocularly, intrapericardially, intraperitoneally,intrapleurally, intraprostatically, intrarectally, intrathecally,intratracheally, intratumorally, intraumbilically, intravaginally,intravenously, intravesicularlly, intravitreally, liposomally, locally,mucosally, parenterally, rectally, subconjunctival, subcutaneously,sublingually, topically, transbuccally, transdermally, vaginally, incrmes, in lipid compositions, via a catheter, via a lavage, viacontinuous infusion, via infusion, via inhalation, via injection, vialocal delivery, or via localized perfusion. In some embodiments, thecomposition is formulated for administration: orally, intravenously, ortopically.

In yet another aspect, the present disclosure provides a method oftreating a disease or disorder comprising administering apharmaceutically effective amount of a compound or composition asdescribed herein. In some embodiments, the disease or disorder is amicrobial infection. In some embodiments, the microbial infection is abacterial infection. In some embodiments, the infection is by a grampositive or gram negative bacteria. In some embodiments, the disease isa bacteria infection by Enterococcus faecalis, Enterococcus faecium,Staphylococcus aureus, Acinetobacter baumannii, Escherichia coli,Acinetobacter calcoaceticus, Staphycococcus epidermidis, Pseudomonasaeruginosa, Klebsiella aerogenes, Candida albicans, Salmonellatyphinurium, Streptococcus pneumoniae, Micrococcus luteus, Bacilluscerues, or Bacillus subtilis. In some embodiments, the disease is abacteria infection by Staphylococcus aureus 503, Staphylococcus aureus209, Staphylococcus aureus RN420, Methicillin-resistant Staphylococcusaureus CCARM 3167, Methicillin-resistant Staphylococcus aureus 371,Methicillin-resistant Staphylococcus aureus CCARM 3506,quinolone-resistant Staphylococcus aureus CCARM 3505,quinolone-resistant Staphylococcus aureus CCARM 3519, Bacillus subtilisKCTC 1021, Bacillus cerues KCTC 1661, Micrococcus luteus KCTC 1056,Streptococcus pneumoniae KCTC 3932, Streptococcus pneumoniae KCTC 5412,Enterococcus faecium 501, Enterococcus faecium KCTC 3122, Enterococcusfaecalis 5613, Enterococcus faecalis KCTC 5191, Enterococcus faecalisKCTC 3511, Staphycococcus epidermidis KCTC 3958, Salmonella typhinuriumKCTC 1926, Acinetobacter calcoaceticus KCTC 2357, Escherichia coli CCARM1358, Escherichia coli KCTC 1682, Pseudomonas aeruginosa KCTC 2004,Pseudomonas aeruginosa KCTC 2742, Klebsiella aerogenes KCTC 2619,Acinetobacter baumannii AB210, or Candida albicans KCTC 7535. In someembodiments, the bacteria is a drug-resistant bacteria. In someembodiments, the method further comprises administering a secondtherapeutic agent. In some embodiments, the second therapeutic agent isan antibiotic. In some embodiments, the second therapeutic agent is atetracycline antibiotic. In some embodiments, the second therapeuticagent is viridicatumtoxin A, viridicatumtoxin B, vancomycin,tetracycline, spirohexaline, minocycline, tigecycline, doxycycline, aβ-lactam antibiotic, an aminoglycoside antibiotic, a sulfonamideantibiotic, a macrolide antibiotic, a glycopeptide antibioitic, anansamycin antibiotic, an oxazolidinone antibiotic, a quinoloneantibiotic, a streptogramin antibiotic, or a lipopeptide antibiotic. Inother embodiments, the microbial infection is a viral infection. In someembodiments, the virus is a poxvirus. In some embodiments, the poxvirusis variola virus, vaccinia virus, or molluscum contagiosum. In someembodiments, the method further comprises administering a secondtherapeutic agent. In some embodiments, the second therapeutic agent isan interferon or antiviral compound. In other embodiments, the diseaseor disorder is cancer. In some embodiments, the cancer is a carcinoma,sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma,or seminoma. In some embodiments, the cancer is of the bladder, blood,bone, brain, breast, central nervous system, cervix, colon, endometrium,esophagus, gall bladder, gastrointestinal tract, genitalia,genitourinary tract, head, kidney, larynx, liver, lung, muscle tissue,neck, oral or nasal mucosa, ovary, pancreas, prostate, skin, spleen,small intestine, large intestine, stomach, testicle, or thyroid. In someembodiments, the method further comprises administering a secondtherapeutic agent. In some embodiments, the second therapeutic agent isa second chemotherapeutic agent, radiotherapy, immunotherapy, orsurgery.

In another aspect, the present disclosure provides a method ofinhibiting the activity of a bacterial ribosome for the treatment of adisease or disorder comprising administering a compound or compositionaccording to any one of claims 1-58. In still another aspect, thepresent disclosure provides a method of inhibiting the activity of abacterial UPP synthase for the treatment of a disease or disordercomprising administering a compound or composition as described herein

In yet another aspect, the present disclosure provides a method ofpreparing a compound of the formula:

wherein: Y₄, Y₅, and Y₆ are each independently hydrogen, alkyl_((C≦8))or substituted alky_((C≦8)); R₁₆ and R₁₆′ are each independentlyhydrogen, alkoxy_((C≦8)), aralkoxy_((C≦12)), substituted alkoxy_((C≦8)),or substituted aralkoxy_((C≦12)); or R₁₆ and R₁₆′ are taken together andare alkoxydiyl_((C≦12)); R₁₂ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), or a substituted version ofany of these groups; or —C(O)OCH₂CH₂Si(CH₃)₃; X₃ is O, NH, S, or CH₂; X₄is hydroxy, amino, mercapto, O, NH, or S provided that when X₄ is O, NH,or S, the bond to which the atom is attached is a double bond andprovide that when the bond to which the atom is attached is a doublebond, then X₄ is O, NH, or S; X₅ is O, NH, or S; R₁₈ is hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; or—X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12)) or substitutedalkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido, carboxy, orcyano, alkenyl_((C≦6)), alkynyl_((C≦6)), heterocycloalkyl_((C≦12)),alkylamino_((C≦8)), dialkylamino_((C≦8)), alkoxy_((C≦8)), or asubstituted version of any of these groups; R₁₉ is hydrogen, amino,carboxy, cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)),alkoxy_((C≦12)), alkylamino_((C≦12)), dialkylamino_((C≦18)),amido_((C≦12)), or a substituted version of any of these groups; or—NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen, alkyl_((C≦8)), orsubstituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8)) or substitutedalkanediyl_((C≦8)); and R₁₄ and R₁₅ are each independently selectedfrom: hydrogen, alkyl_((C≦12)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦12)), or a substituted version of any of these groups; or R₁₄and R₁₅ are taken together and are alkanediyl_((C≦8)),alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or a substituted version ofany of these groups; and R₂₀ and R₂₁ are each independently hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; or asalt, tautomer, or optical isomer thereof; comprising reacting acompound of the formula:

wherein: Y₄, Y₅, Y₆, R₁₆, R₁₆′, X₅, R₁₈, R₁₉, and R₂₀ are as definedabove; and X₄ is O, NH, or S; with a compound of the formula:

wherein: X₃ and R₁₇ are as defined above; R₂₁ is hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; and R₂₂is aryl_((C≦12)), aralkyl_((C≦12)), or a substituted version of eitherof these groups; in the presence of a base. In some embodiments, Y₄, Y₅,and Y₆ are alkyl_((C≦6)). In some embodiments, Y₄, Y₅, and Y₆ aremethyl. In some embodiments, R₁₆ and R₁₆′ are alkoxy_((C≦8)) orsubstituted alkoxy_((C≦8)). In some embodiments, R₁₆ and R₁₆′ aremethoxy. In some embodiments, R₁₇ is hydrogen, amino, halo, hydroxy;alkylamino_((C≦12)), substituted alkylamino_((C≦12)),dialkylamino_((C≦18)), substituted dialkylamino_((C≦18)), or—C(O)OCH₂CH₂Si(CH₃)₃; In some embodiments, R₁₇ is amino, hydroxy, orhalo. In other embodiments, R₁₇ is dimethylamino or2-aminoethylmethylamino. In some embodiments, X₃ and X₅ are O. In someembodiments, X₄ is hydroxy. In other embodiments, X₄ is O. In someembodiments, R₁₈ is hydrogen, alkyl_((C≦8)), substituted alkyl_((C≦8)),or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12)) or substitutedalkanediyl_((C≦12)) and R₁₁ is hydroxy, amino,heterocycloalkyl_((C≦12)), substituted heterocycloalkyl_((C≦12)),alkylamino_((C≦8)), or substituted alkylamino_((C≦8)), In someembodiments, R₁₈ is hydrogen. In some embodiments, R₁₉ is hydrogen,amino, halo, hydroxy, alkylamino_((C≦12)), substitutedalkylamino_((C≦12)), amido_((C≦12)), substituted amido_((C≦12)), or—NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen, R₁₃ isalkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅are each independently hydrogen, alkyl_((C≦12)), substitutedalkyl_((C≦12)), aryl_((C≦12)), substituted aryl_((C≦12)),aralkyl_((C≦12)), substituted aralkyl_((C≦12)), acyl_((C≦12)), orsubstituted acyl_((C≦12)). In some embodiments, R₁₉ is hydrogen, halo,amido_((C≦12)), substituted amido_((C≦12)), or —NHC(O)CH₂—NHR₁₅ whereinR₁₅ is hydrogen, alkyl_((C≦12)), or substituted alkyl_((C≦12)). In someembodiments, R₂₀ is hydrogen, alkyl_((C≦6)), or substitutedalkyl_((C≦6)). In some embodiments, R₂₀ is methyl. In some embodiments,R₂₁ is aralkyl_((C≦12)) or substituted aralkyl_((C≦12)). In someembodiments, R₂₁ is benzyl. In some embodiments, R₂₂ is aryl_((C≦12)) orsubstituted aryl_((C≦12)). In some embodiments, R₂₂ is phenyl. In otherembodiments, R₁₇ is not hydrogen or —C(O)OCH₂CH₂Si(CH₃)₃, R₁₈ is notbenzyl, R₁₉ are not hydrogen, and R₂₀ is not methyl. In someembodiments, the base is an alkoxide_((C≦18)). In some embodiments, thealkoxide_((C≦18)) is t-butoxide. In some embodiments, the methodcomprises adding from about 1.0 to about 2.0 equivalents of baserelative to the compound of formula VII. In some embodiments, the methodcomprises adding from about 1.0 to about 1.5 equivalents of base. Insome embodiments, the method comprises adding about 1.2 equivalents ofbase. In some embodiments, the method comprises adding from about 0.9 toabout 2.0 equivalents of the compound of formula VIII relative to thecompound of formula VII. In some embodiments, the method comprisesadding from about 1.0 to about 1.5 equivalents of the compound offormula VIII. In some embodiments, the method comprises adding about 1.1equivalents of the compound of formula VIII. In some embodiments, themethod further comprises a solvent. In some embodiments, the solvent isan organic solvent. In some embodiments, the solvent is anarene_((C≦12)). In some embodiments, the solvent is toluene. In someembodiments, the reaction comprises running the reaction at atemperature from about 0° C. to about 50° C. In some embodiments, thetemperature is from about 20° C. to about 35° C. In some embodiments,the temperature is about 25° C. In some embodiments, the temperature isabout room temperature. In some embodiments, the reaction is run for atime period from about 5 minutes to about 2 hours. In some embodiments,the time period is from about 10 minutes to about 45 minutes. In someembodiments, the time period is about 15 minutes. In some embodiments,the reaction results in a yield of greater than about 50%. In someembodiments, the yield is greater than about 75%. In some embodiments,the yield is greater than about 90%. In some embodiments, the reactionproduces a diastereomeric ratio of greater than about 1:1. In someembodiments, the diastereomeric ratio is greater than about 1.75:1. Insome embodiments, the diastereomeric ratio is about 2:1.

In some embodiments, the method further comprises forming a compound ofthe formula:

wherein: Y₄, Y₅, and Y₆ are each independently hydrogen, alkyl_((C≦8))or substituted alky_((C≦8)); R₁₆ and R₁₆′ are each independentlyhydrogen, alkoxy_((C≦8)), aralkoxy_((C≦12)), substituted alkoxy_((C≦8)),or substituted aralkoxy_((C≦12)); or R₁₆ and R₁₆′ are taken together andare alkoxydiyl_((C≦12)); R₁₇ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), or a substituted version ofany of these groups; or —C(O)OCH₂CH₂Si(CH₃)₃; X₃ is O, NH, S, or CH₂; X₄is O, NH, or S; X₅ is O, NH, or S; R₁₈ is hydrogen, alkyl_((C≦8)),alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or asubstituted version of any of these groups; or —X₂—R₁₁, wherein: X₂ isalkanediyl_((C≦12)) or substituted alkanediyl_((C≦12)); and R₁₁ ishydroxy, amino, azido, carboxy, or cyano, alkenyl_((C≦6)),alkynyl_((C≦6)), heterocycloalkyl_((C≦12)), alkylamino_((C≦8)),dialkylamino_((C≦8)), alkoxy_((C≦8)), or a substituted version of any ofthese groups; R₁₉ is hydrogen, amino, carboxy, cyano, halo, hydroxy,nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; and R₂₀ and R₂₁ are eachindependently hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦8)), or a substituted version of any of thesegroups; or a salt, tautomer, or optical isomer thereof, comprisingreacting a compound of the formula:

wherein the variables are as defined above; with a metal catalyst and anoxidizing agent. In some embodiments, R₁₇ is not hydrogen, R₁₈ is notbenzyl, R₁₉ is not hydrogen, and R₂₀ is not methyl. In some embodiments,the metal catalyst is a nickel(II) salt. In some embodiments, thenickel(II) salt is Ni(acac)₂. In some embodiments, the reactioncomprises adding from about 0.01 to about 1.0 equivalents of the metalcatalyst relative to the compound of formula X. In some embodiments, thereaction comprises adding from about 0.1 to about 0.5 equivalents of themetal catalyst. In some embodiments, the reaction comprises adding about0.2 equivalents of the metal catalyst. In some embodiments, theoxidizing agent is a dioxirane compound. In some embodiments, theoxidizing agent is dimethyldioxirane (DMDO). In some embodiments, thereaction comprises adding from about 1.5 equivalents to about 10.0equivalents of dimethyldioxirane relative to the compound of formula X.In some embodiments, the reaction comprises adding from about 4.0equivalents to about 6.0 equivalents of dimethyldioxirane. In someembodiments, the reaction comprises adding about 5.1 equivalents ofdimethyldioxirane. In some embodiments, the method further comprisesadding additional dimethyldioxirane every two hours during the reaction.In some embodiments, the additional dimethyldioxirane is about 1.5equivalents relative to the compound of formula X. In some embodiments,the method further comprises a solvent. In some embodiments, the solventis an organic solvent. In some embodiments, the solvent is ahaloalkane_((C≦12)). In some embodiments, the solvent isdichloromethane. In some embodiments, the reaction comprises running thereaction at a temperature from about −90° C. to about −40° C. In someembodiments, the temperature is from about −80° C. to about −60° C. Insome embodiments, the temperature is about −78° C. In some embodiments,the method further comprises allowing the reaction to warm to atemperature from about −80° C. to about −30° C. In some embodiments, thetemperature is about −60° C. In some embodiments, the reaction is runfor a time period from about 3 hours to about 12 hours. In someembodiments, the time period is from about 5 hours to about 8 hours. Insome embodiments, the time period is about 6.5 hours. In someembodiments, the starting material is recovered after the time periodand subject to the reaction conditions again. In some embodiments, thereaction results in a yield of greater than about 35% based uponrecovered starting material. In some embodiments, the yield is greaterthan about 50%. In some embodiments, the yield is greater than about60%. In some embodiments, the reaction produces a diastereomeric ratioof greater than about 1:1. In some embodiments, the diastereomeric ratiois greater than about 1.75:1. In some embodiments, the diastereomericratio is about 2:1.

In some embodiments, the reaction further comprises reacting a compoundof formula IX with a reducing agent to form a compound of the formula:

wherein: Y₄, Y₅, and Y₆ are each independently hydrogen, alkyl_((C≦8))or substituted alky_((C≦8)); R₁₆ and R₁₆′ are each independentlyhydrogen, alkoxy_((C≦8)), aralkoxy_((C≦12)), substituted alkoxy_((C≦8)),or substituted aralkoxy_((C≦12)); or R₁₆ and R₁₆′ are taken together andare alkoxydiyl_((C≦12)); R₁₇ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), or a substituted version ofany of these groups; or —C(O)OCH₂CH₂Si(CH₃)₃; X₃ is O, NH, S, or CH₂; X₄is O, NH, or S; R₁₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; R₁₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; R₂₀ and R₂₁ are eachindependently hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦8)), or a substituted version of any of thesegroups; and R₂₃ is hydroxy, alkoxy_((C≦12)), or substitutedalkoxy_((C≦12)); or a salt, tautomer, or optical isomer thereof. In someembodiments, the reducing agent is a soft hydride donor. In someembodiments, the reducing agent is a metal borohydride reagent. In someembodiments, the reducing agent is sodium cyanoborohydride. In someembodiments, the reaction comprises adding from about 5.0 equivalents toabout 25.0 equivalents of reducing agent relative to the compound offormula IX. In some embodiments, the reaction comprises adding fromabout 8.0 equivalents to about 12.0 equivalents of reducing agent. Insome embodiments, the reaction comprises adding about 10.0 equivalentsof reducing agent. In some embodiments, the method further comprises asolvent. In some embodiments, the solvent is an organic solvent. In someembodiments, the solvent is a ether_((C≦12)). In some embodiments, thesolvent is tetrahydrofuran. In some embodiments, the reaction comprisesrunning the reaction at a temperature from about −90° C. to about −40°C. In some embodiments, the temperature is from about −80° C. to about−60° C. In some embodiments, the temperature is about 78° C. In someembodiments, the method further comprises allowing the reaction to warmto a temperature from about −80° C. to about −30° C. In someembodiments, the temperature is about −60° C. In some embodiments, thereaction is run for a time period from about −30 minutes to about 6hours. In some embodiments, the time period is from about 1 hour toabout 3 hours. In some embodiments, the time period is about 1.5 hours.In some embodiments, the reaction results in a yield of greater thanabout 20%. In some embodiments, the yield is greater than about 25%. Insome embodiments, the yield is greater than about 35%. In someembodiments, the reaction produces a diastereomeric ratio of greaterthan about 1:1. In some embodiments, the diastereomeric ratio is greaterthan about 1.75:1. In some embodiments, the diastereomeric ratio isabout 2:1.

In some embodiments, one or more steps of the reaction further comprisespurifying the reaction in a purification step. In some embodiments, thepurification method is chromatography. In some embodiments, thepurification method is column chromatography or high performance liquidchromatography.

It is contemplated that any method or composition described herein canbe implemented with respect to any other method or composition describedherein. For example, an aldehyde synthesized by one method may be usedin the preparation of a final compound according to a different method.

The use of the word “a” or “an” when used in conjunction with the term“comprising” in the claims and/or the specification may mean “one,” butit is also consistent with the meaning of “one or more,” “at least one,”and “one or more than one.” The word “about” means plus or minus 5% ofthe stated number.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and the specificexamples, while indicating specific embodiments of the invention, aregiven by way of illustration only, since various changes andmodifications within the spirit and scope of the invention will becomeapparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE FIGURES

The following drawings form part of the present specification and areincluded to further demonstrate certain aspects of the presentinvention. The invention may be better understood by reference to one ormore of these drawings in combination with the detailed.

FIGS. 1A & B—Structures of bacterial tetracyclines and designed analogs(1A) and fungal tetracyclines (1B)

FIG. 2—Structures of Viridicatumtoxin B and A (1 and 2, respectively),original proposed structure of Viridicatumtoxin B, and spirohexaline.

FIG. 3—Biosynthetic Pathway of Viridicatumtoxin A production in vivo.

FIG. 4—Structures of Viridicatumtoxin B and A (1 and 2, respectively)and analogs (V2-V6).

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The present disclosure relates to a series of novel analogs ofviridicatumtoxin and an improved synthetic pathway to obtainviridicatumtoxin and its analogs. In some aspects, the analogs of thefungal secondary metabolites viridicatumtoxin A (2) and B (1) (FIG. 4)are useful as potent antibiotics against a variety of Gram-positive andcertain Gram-negative bacterial strains. In the present disclosure, acollection of viridicatumtoxin analogs (V2V6, FIG. 4) are synthesizedand their antibiotic profile is evaluated. These and other aspects ofthe disclosure are described in greater detail below.

I. COMPOUNDS AND FORMULATIONS THEREOF

In one aspect, the present invention provides compounds of the formula:

wherein: X₁ is absent such that atoms 16 and 17 are only connected bythe shown single bond, a covalent bond such that a double bond is formedbetween atoms 16 and 17, —O—, alkanediyl_((C≦8)), or substitutedalkanediyl_((C≦8)); Y₁, Y₂, and Y₃ are each independently alkyl_((C≦12))or substituted alkyl_((C≦12)); R₁ is hydrogen, hydroxy, or oxo, providedthat when R₁ is oxo, the bond between R₁ and atom number 5 is a doublebond and when the bond between R₁ and atom number 5 is a double bondthen R₁ is oxo, alkoxy_((C≦8)), or substituted alkoxy_((C≦8)); R₂ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₃, R₆, R₂, and R₁₀ are each independently selected from: hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; R₄ and R₅are each independently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.

Additionally, the compounds provided by the present disclosure areshown, for example, above in the summary of the invention section and inthe examples and claims below. They may be made using the methodsoutlined in the Examples section. Viridicatumtoxin and its derivativescan be synthesized according to the methods described, for example, inthe Examples section below. These methods can be further modified andoptimized using the principles and techniques of organic chemistry asapplied by a person skilled in the art. Such principles and techniquesare taught, for example, in March's Advanced Organic Chemistry:Reactions, Mechanisms, and Structure (2007), which is incorporated byreference herein.

Viridicatumtoxin and its derivatives of the disclosure may contain oneor more asymmetrically-substituted carbon or nitrogen atoms, and may beisolated in optically active or racemic form. Thus, all chiral,diastereomeric, racemic form, epimeric form, and all geometric isomericforms of a chemical formula are intended, unless the specificstereochemistry or isomeric form is specifically indicated. Compoundsmay occur as racemates and racemic mixtures, single enantiomers,diastereomeric mixtures and individual diastereomers. In someembodiments, a single diastereomer is obtained. The chiral centers ofthe compounds of the present invention can have the S or the Rconfiguration.

Chemical formulas used to represent viridicatumtoxin and its derivativesof the disclosure will typically only show one of possibly severaldifferent tautomers. For example, many types of ketone groups are knownto exist in equilibrium with corresponding enol groups. Similarly, manytypes of imine groups exist in equilibrium with enamine groups.Regardless of which tautomer is depicted for a given compound, andregardless of which one is most prevalent, all tautomers of a givenchemical formula are intended.

Viridicatumtoxin and its derivatives of the disclosure may also have theadvantage that they may be more efficacious than, be less toxic than, belonger acting than, be more potent than, produce fewer side effectsthan, be more easily absorbed than, and/or have a better pharmacokineticprofile (e.g., higher oral bioavailability and/or lower clearance) than,and/or have other useful pharmacological, physical, or chemicalproperties over, compounds known in the prior art, whether for use inthe indications stated herein or otherwise.

In addition, atoms making up viridicatumtoxin and its derivatives of thepresent disclosure are intended to include all isotopic forms of suchatoms. Isotopes, as used herein, include those atoms having the sameatomic number but different mass numbers. By way of general example andwithout limitation, isotopes of hydrogen include tritium and deuterium,and isotopes of carbon include ¹³C and ¹⁴C.

Viridicatumtoxin and its derivatives of the present disclosure may alsoexist in prodrug form. Since prodrugs are known to enhance numerousdesirable qualities of pharmaceuticals (e.g., solubility,bioavailability, manufacturing, etc.), the compounds employed in somemethods of the disclosure may, if desired, be delivered in prodrug form.Thus, the invention contemplates prodrugs of compounds of the presentinvention as well as methods of delivering prodrugs. Prodrugs ofviridicatumtoxin and its derivatives employed in the disclosure may beprepared by modifying functional groups present in the compound in sucha way that the modifications are cleaved, either in routine manipulationor in vivo, to the parent compound. Accordingly, prodrugs include, forexample, compounds described herein in which a hydroxy, amino, orcarboxy group is bonded to any group that, when the prodrug isadministered to a subject, cleaves to form a hydroxy, amino, orcarboxylic acid, respectively.

It should be recognized that the particular anion or cation forming apart of any salt form of a compound provided herein is not critical, solong as the salt, as a whole, is pharmacologically acceptable.Additional examples of pharmaceutically acceptable salts and theirmethods of preparation and use are presented in Handbook ofPharmaceutical Salts: Properties, and Use (2002), which is incorporatedherein by reference.

Those skilled in the art of organic chemistry will appreciate that manyorganic compounds can form complexes with solvents in which they arereacted or from which they are precipitated or crystallized. Thesecomplexes are known as “solvates.” For example, a complex with water isknown as a “hydrate.” Solvates of viridicatumtoxin and its derivativesprovided herein are within the scope of the invention. It will also beappreciated by those skilled in organic chemistry that many organiccompounds can exist in more than one crystalline form. For example,crystalline form may vary from solvate to solvate. Thus, all crystallineforms of viridicatumtoxin and its derivatives or the pharmaceuticallyacceptable solvates thereof are within the scope of the presentinvention.

B. Formulations

In some embodiments of the present disclosure, the compounds areincluded a pharmaceutical formulation. Materials for use in thepreparation of microspheres and/or microcapsules are, e.g.,biodegradable/bioerodible polymers such as polygalactin, poly-(isobutylcyanoacrylate), poly(2-hydroxyethyl-L-glutam-nine) and, poly(lacticacid). Biocompatible carriers that may be used when formulating acontrolled release parenteral formulation are carbohydrates (e.g.,dextrans), proteins (e.g., albumin), lipoproteins, or antibodies.Materials for use in implants can be non-biodegradable (e.g.,polydimethyl siloxane) or biodegradable (e.g., poly(caprolactone),poly(lactic acid), poly(glycolic acid) or poly(ortho esters) orcombinations thereof).

Formulations for oral use include tablets containing the activeingredient(s) (e.g., viridicatumtoxin and its derivatives) in a mixturewith non-toxic pharmaceutically acceptable excipients. Such formulationsare known to the skilled artisan. Excipients may be, for example, inertdiluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol,microcrystalline cellulose, starches including potato starch, calciumcarbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate,or sodium phosphate); granulating and disintegrating agents (e.g.,cellulose derivatives including microcrystalline cellulose, starchesincluding potato starch, croscarmellose sodium, alginates, or alginicacid); binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginicacid, sodium alginate, gelatin, starch, pregelatinized starch,microcrystalline cellulose, magnesium aluminum silicate,carboxymethylcellulose sodium, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethyleneglycol); and lubricating agents, glidants, and antiadhesives (e.g.,magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenatedvegetable oils, or talc). Other pharmaceutically acceptable excipientscan be colorants, flavoring agents, plasticizers, humectants, bufferingagents, and the like.

The tablets may be uncoated or they may be coated by known techniques,optionally to delay disintegration and absorption in thegastrointestinal tract and thereby providing a sustained action over alonger period. The coating may be adapted to release the active drug ina predetermined pattern (e.g., in order to achieve a controlled releaseformulation) or it may be adapted not to release the active drug untilafter passage of the stomach (enteric coating). The coating may be asugar coating, a film coating (e.g., based on hydroxypropylmethylcellulose, methylcellulose, methyl hydroxyethylcellulose,hydroxypropylcellulose, carboxymethylcellulose, acrylate copolymers,polyethylene glycols and/or polyvinylpyrrolidone), or an enteric coating(e.g., based on methacrylic acid copolymer, cellulose acetate phthalate,hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcelluloseacetate succinate, polyvinyl acetate phthalate, shellac, and/orethylcellulose). Furthermore, a time delay material, such as, e.g.,glyceryl monostearate or glyceryl distearate may be employed.

II. MICROBIAL INFECTIONS

A. Bacterial Infections

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection. While humans containnumerous different bacteria on and inside their bodies, an imbalance inbacterial levels or the introduction of pathogenic bacteria can cause asymptomatic bacterial infection. Pathogenic bacteria cause a variety ofdifferent diseases including but not limited to numerous foodborneillness, typhoid fever, tuberculosis, pneumonia, syphilis, and leprosy.

Additionally, different bacteria have a wide range of interactions withbody and those interactions can modulate ability of the bacteria tocause an infection. For example, bacteria can be conditionallypathogenic such that they only cause an infection under specificconditions. For example, Staphylococcus and Streptococcus bacteria existin the normal human bacterial biome, but these bacteria when they areallowed to colonize other parts of the body causing a skin infection,pneumonia, or sepsis. Other bacteria are known as opportunisticpathogens and only cause diseases in a patient with a weakened immunesystem or another disease or disorder.

Bacteria can also be intracellular pathogens which can grow andreproduce within the cells of the host organism. Such bacteria can bedivided into two major categories as either obligate intracellularparasites or facultative intracellular parasites. Obligate intracellularparasites require the host cell in order to reproduce and include suchbacteria as but are not limited to Chlamydophila, Rickettsia, andEhrlichia which are known to cause pneumonia, urinary tract infections,typhus, and Rocky Mountain spotted fever. Facultative intracellularparasites can reproduce either intracellular or extracellular. Somenon-limiting examples of facultative intracellular parasites includeSalmonella, Listeria, Legionella, Mycobacterium, and Brucella which areknown to cause food poisoning, typhoid fever, sepsis, meningitis,Legionnaire's disease, tuberculosis, leprosy, and brucellosis.

Finally, bacterial infections could be targeted to a specific locationin or on the body. For example, bacteria could be harmless if onlyexposed to the specific organs, but when it comes in contact with aspecific organ or tissue, the bacteria can begin replicating and cause abacterial infection.

In particular, the inventors contemplate treatment of bacterialinfections, including those caused by Staphyloccoccus aureus. S. aureusis a major human pathogen, causing a wide variety of illnesses rangingfrom mild skin and soft tissue infections and food poisoning tolife-threatening illnesses such as deep post-surgical infections,septicaemia, endocarditis, necrotizing pneumonia, and toxic shocksyndrome. These organisms have a remarkable ability to accumulateadditional antibiotic resistance determinants, resulting in theformation of multiply-drug-resistant strains.

Methicillin, being the first semi-synthetic penicillin to be developed,was introduced in 1959 to overcome the problem of penicillin-resistantS. aureus due to β-lactamase (penicillinase) production (Livermore,2000). However, methicillin-resistant S. aureus (MRSA) strains wereidentified soon after the introduction of methicillin (Barber, 1961;Jevons, 1961). MRSA have acquired and integrated into their genome a 21-to 67-kb mobile genetic element, termed the staphylococcal cassettechromosome mec (SCCmec) that harbors the methicillin resistance (mecA)gene and other antibiotic resistance determinants (Ito et al., 2001; Itoet al., 2004; Ma et al., 2002). The mecA gene encodes an alteredadditional low affinity penicillin-binding protein (PBP2a) that confersbroad resistance to all penicillin-related compounds includingcephalosporins and carbapenems that are currently some of the mostpotent broad-spectrum drugs available (Hackbarth & Chambers, 1989).Since their first identification, strains of MRSA have spread and becomeestablished as major nosocomial (hospital-acquired (HA)-MRSA) pathogensworldwide (Ayliffe, 1997; Crossley et al., 1979; Panlilio et al., 1992;Voss et al., 1994). These organisms have evolved and emerged as a majorcause of community-acquired infections (CA-MRSA) in healthy individualslacking traditional risk factors for infection, and are causingcommunity-outbreaks, which pose a significant threat to public health.

i. Gram Positive Bacteria

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection by a gram positivebacteria. Gram positive bacteria contain a thick peptidoglycan layerwithin the cell wall which prevents the bacteria from releasing thestain when dyed with crystal violet. Without being bound by theory, thegram positive bacteria are often more susceptible to antibiotics.Generally, gram positive bacteria, in addition to the thickpeptidoglycan layer, also comprise a lipid monolayer and containteichoic acids which react with lipids to form lipoteichoic acids thatcan act as a chelating agent. Additionally, in gram positive bacteria,the peptidoglycan layer is outer surface of the bacteria. Many grampositive bacteria have been known to cause disease including, but arenot limited to, Streptococcus, Straphylococcus, Corynebacterium,Enterococcus, Listeria, Bacillus, Clostridium, Rathybacter, Leifsonia,and Clavibacter.

ii. Gram Negative Bacteria

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection by a gram negativebacteria. Gram negative bacteria do not retain the crystal violet stainafter washing with alcohol. Gram negative bacteria, on the other hand,have a thin peptidoglycan layer with an outer membrane oflipopolysaccharides and phospholipids as well as a space between thepeptidoglycan and the outer cell membrane called the periplasmic space.Gram negative bacterial generally do not have teichoic acids orlipoteichoic acids in their outer coating. Generally, gram negativebacteria also release some endotoxin and contain prions which act asmolecular transport units for specific compounds. Most bacteria are gramnegative. Some non-limiting examples of gram negative bacteria includeBordetella, Borrelia, Burcelia, Campylobacteria, Escherichia,Francisella, Haemophilus, Helicobacter, Legionella, Leptospira,Neisseria, Pseudomonas, Rickettsia, Salmonella, Shigella, Treponema,Vibrio, and Yersinia.

Iii. Gram Indeterminate Bacteria

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection by a gramindeterminate bacteria. Gram indeterminate bacteria do not full stain orpartially stain when exposed to crystal violet. Without being bound bytheory, a gram indeterminate bacteria may exhibit some of the propertiesof the gram positive and gram negative bacteria. A non-limiting exampleof a gram indeterminate bacteria include mycobacterium tuberculosis ormycobacterium leprae.

B. Viral Infections

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a viral infection. Similarly, virus can alsoexist in pathogenic form which can lead to human diseases. Viralinfections are typically not treated directly but rather symptomaticallysince virus often have a self-limiting life cycle. Viral infections canalso be more difficult to diagnosis than a bacterial infection sinceviral infections often do result in the concombinent increase in whiteblood cell counts. Some non-limiting examples of pathogenic virusinclude influenza virus, smallpox, BK virus, JC virus, humanpapillomavirus, adenovirus, herpes simplex type 1, herpes simplex type2, varicella-zoster virus, Epstein barr virus, human cytomegalovirus,human herpesvirus type 8, Norwalk virus, human bocavirus, rubella virus,hepatitis E virus, hepatitis B virus, human immunodeficiency virus(HIV), Ebola virus, rabies virus, rotavirus, and hepatitis D virus.

III. HYPERPROLIFERATIVE DISEASES

A. Cancer and Other Hyperproliferative Disease

While hyperproliferative diseases can be associated with any diseasewhich causes a cell to begin to reproduce uncontrollably, theprototypical example is cancer. One of the key elements of cancer isthat the cell's normal apoptotic cycle is interrupted and thus agentsthat interrupt the growth of the cells are important as therapeuticagents for treating these diseases. In this disclosure, theviridicatumtoxin derivatives may be used to lead to decreased cellcounts and as such can potentially be used to treat a variety of typesof cancer lines. In various aspects, it is anticipated that theviridicatumtoxin derivatives of the present disclosure may be used totreat virtually any malignancy.

Cancer cells that may be treated with the compounds of the presentdisclosure include but are not limited to cells from the bladder, blood,bone, bone marrow, brain, breast, colon, esophagus, gastrointestine,gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate,skin, stomach, pancreas, testis, tongue, cervix, or uterus. In addition,the cancer may specifically be of the following histological type,though it is not limited to these: neoplasm, malignant; carcinoma;carcinoma, undifferentiated; giant and spindle cell carcinoma; smallcell carcinoma; papillary carcinoma; squamous cell carcinoma;lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma;transitional cell carcinoma; papillary transitional cell carcinoma;adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma;hepatocellular carcinoma; combined hepatocellular carcinoma andcholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma;adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposiscoli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolaradenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma;acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clearcell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma;papillary and follicular adenocarcinoma; nonencapsulating sclerosingcarcinoma; adrenal cortical carcinoma; endometroid carcinoma; skinappendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma;ceruminous adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma;papillary cystadenocarcinoma; papillary serous cystadenocarcinoma;mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cellcarcinoma; infiltrating duct carcinoma; medullary carcinoma; lobularcarcinoma; inflammatory carcinoma; paget's disease, mammary; acinar cellcarcinoma; adenosquamous carcinoma; adenocarcinoma w/squamousmetaplasia; thymoma, malignant; ovarian stromal tumor, malignant;thecoma, malignant; granulosa cell tumor, malignant; androblastoma,malignant; sertoli cell carcinoma; leydig cell tumor, malignant; lipidcell tumor, malignant; paraganglioma, malignant; extra-mammaryparaganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignantmelanoma; amelanotic melanoma; superficial spreading melanoma; maligmelanoma in giant pigmented nevus; epithelioid cell melanoma; bluenevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma,malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma;embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma;mixed tumor, malignant; mullerian mixed tumor; nephroblastoma;hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; brenner tumor,malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma,malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant;struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant;hemangiosarcoma; hemangioendothelioma, malignant; kaposi's sarcoma;hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma;juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant;mesenchymal chondrosarcoma; giant cell tumor of bone; ewing's sarcoma;odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma,malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma;glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma;fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma;oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma;ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactoryneurogenic tumor; meningioma, malignant; neurofibrosarcoma;neurilemmoma, malignant; granular cell tumor, malignant; malignantlymphoma; hodgkin's disease; hodgkin's; paragranuloma; malignantlymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse;malignant lymphoma, follicular; mycosis fungoides; other specifiednon-hodgkin's lymphomas; malignant histiocytosis; multiple myeloma; mastcell sarcoma; immunoproliferative small intestinal disease; leukemia;lymphoid leukemia; plasma cell leukemia; erythroleukemia; lymphosarcomacell leukemia; myeloid leukemia; basophilic leukemia; eosinophilicleukemia; monocytic leukemia; mast cell leukemia; megakaryoblasticleukemia; myeloid sarcoma; and hairy cell leukemia. In certain aspects,the tumor may comprise an osteosarcoma, angiosarcoma, rhabdosarcoma,leiomyosarcoma, Ewing sarcoma, glioblastoma, neuroblastoma, or leukemia.

III. THERAPIES

A. Pharmaceutical Formulations and Routes of Administration

Where clinical applications are contemplated, it will be necessary toprepare pharmaceutical compositions in a form appropriate for theintended application. In some embodiments, such formulation with thecompounds of the present disclosure is contemplated. Generally, thiswill entail preparing compositions that are essentially free ofpyrogens, as well as other impurities that could be harmful to humans oranimals.

One will generally desire to employ appropriate salts and buffers torender delivery vectors stable and allow for uptake by target cells.Buffers also will be employed when recombinant cells are introduced intoa patient. Aqueous compositions of the present invention comprise aneffective amount of the vector to cells, dissolved or dispersed in apharmaceutically acceptable carrier or aqueous medium. Such compositionsalso are referred to as inocula. The phrase “pharmaceutically orpharmacologically acceptable” refers to molecular entities andcompositions that do not produce adverse, allergic, or other untowardreactions when administered to an animal or a human. As used herein,“pharmaceutically acceptable carrier” includes any and all solvents,dispersion media, coatings, antibacterial and antifungal agents,isotonic and absorption delaying agents and the like. The use of suchmedia and agents for pharmaceutically active substances is well known inthe art. Except insofar as any conventional media or agent isincompatible with the vectors or cells of the present invention, its usein therapeutic compositions is contemplated. Supplementary activeingredients also can be incorporated into the compositions.

The active compositions of the present invention may include classicpharmaceutical preparations. Administration of these compositionsaccording to the present invention will be via any common route so longas the target tissue is available via that route. Such routes includeoral, nasal, buccal, rectal, vaginal or topical route. Alternatively,administration may be by orthotopic, intradermal, subcutaneous,intramuscular, intratumoral, intraperitoneal, or intravenous injection.Such compositions would normally be administered as pharmaceuticallyacceptable compositions, described supra.

The active compounds may also be administered parenterally orintraperitoneally. Solutions of the active compounds as free base orpharmacologically acceptable salts can be prepared in water suitablymixed with a surfactant, such as hydroxypropylcellulose. Dispersions canalso be prepared in glycerol, liquid polyethylene glycols, and mixturesthereof and in oils. Under ordinary conditions of storage and use, thesepreparations contain a preservative to prevent the growth ofmicroorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms, such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), suitable mixtures thereof,and vegetable oils. The proper fluidity can be maintained, for example,by the use of a coating, such as lecithin, by the maintenance of therequired particle size in the case of dispersion and by the use ofsurfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many cases, it will be preferable to include isotonicagents, for example, sugars or sodium chloride. Prolonged absorption ofthe injectable compositions can be brought about by the use in thecompositions of agents delaying absorption, for example, aluminummonostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompounds in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

As used herein, “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents and the like. The use ofsuch media and agents for pharmaceutical active substances is well knownin the art. Except insofar as any conventional media or agent isincompatible with the active ingredient, its use in the therapeuticcompositions is contemplated. Supplementary active ingredients can alsobe incorporated into the compositions.

For oral administration viridicatumtoxin and its derivatives may beincorporated with excipients and used in the form of non-ingestiblemouthwashes and dentifrices. A mouthwash may be prepared incorporatingthe active ingredient in the required amount in an appropriate solvent,such as a sodium borate solution (Dobell's Solution). Alternatively, theactive ingredient may be incorporated into an antiseptic wash containingsodium borate, glycerin and potassium bicarbonate. The active ingredientmay also be dispersed in dentifrices, including: gels, pastes, powdersand slurries. The active ingredient may be added in a therapeuticallyeffective amount to a paste dentifrice that may include water, binders,abrasives, flavoring agents, foaming agents, and humectants.

The compositions of the present disclosure may be formulated in aneutral or salt form. Pharmaceutically-acceptable salts include the acidaddition salts (formed with the free amino groups of the protein) andwhich are formed with inorganic acids such as, for example, hydrochloricor phosphoric acids, or such organic acids as acetic, oxalic, tartaric,mandelic, and the like. Salts formed with the free carboxyl groups canalso be derived from inorganic bases such as, for example, sodium,potassium, ammonium, calcium, or ferric hydroxides, and such organicbases as isopropylamine, trimethylamine, histidine, procaine and thelike.

Upon formulation, solutions will be administered in a manner compatiblewith the dosage formulation and in such amount as is therapeuticallyeffective. The formulations are easily administered in a variety ofdosage forms such as injectable solutions, drug release capsules and thelike. For parenteral administration in an aqueous solution, for example,the solution should be suitably buffered if necessary and the liquiddiluent first rendered isotonic with sufficient saline or glucose. Theseparticular aqueous solutions are especially suitable for intravenous,intramuscular, subcutaneous and intraperitoneal administration. In thisconnection, sterile aqueous media which can be employed will be known tothose of skill in the art in light of the present disclosure. Forexample, one dosage could be dissolved in 1 ml of isotonic NaCl solutionand either added to 1000 ml of hypodermoclysis fluid or injected at theproposed site of infusion, (see for example, “Remington's PharmaceuticalSciences,” 15th Edition, pages 1035-1038 and 1570-1580). Some variationin dosage will necessarily occur depending on the condition of thesubject being treated. The person responsible for administration will,in any event, determine the appropriate dose for the individual subject.Moreover, for human administration, preparations should meet sterility,pyrogenicity, general safety and purity standards as required by FDAOffice of Biologics standards.

B. Methods of Treatment

In particular, the compositions that may be used in treating microbialinfections and cancer in a subject (e.g., a human subject) are disclosedherein. The compositions described above are preferably administered toa mammal (e.g., rodent, human, non-human primates, canine, bovine,ovine, equine, feline, etc.) in an effective amount, that is, an amountcapable of producing a desirable result in a treated subject (e.g.,causing apoptosis of cancerous cells or killing bacterial cells).Toxicity and therapeutic efficacy of the compositions utilized inmethods of the invention can be determined by standard pharmaceuticalprocedures. As is well known in the medical and veterinary arts, dosagefor any one animal depends on many factors, including the subject'ssize, body surface area, body weight, age, the particular composition tobe administered, time and route of administration, general health, theclinical symptoms of the infection or cancer and other drugs beingadministered concurrently. A composition as described herein istypically administered at a dosage that inhibits the growth orproliferation of a bacterial cell, inhibits the growth of a biofilm, orinduces death of cancerous cells (e.g., induces apoptosis of a cancercell), as assayed by identifying a reduction in hematological parameters(Complete blood count (CBC)), or cancer cell growth or proliferation. Insome embodiments, in the experiments described herein and based upondosing of other tetracycline compounds, the amount of theviridicatumtoxin derivatives used to inhibit bacterial growth, treat aviral infection, or induce apoptosis of the cancer cells is calculatedto be from about 0.01 mg to about 10,000 mg/day. In some embodiments,the amount is from about 1 mg to about 1,000 mg/day. In someembodiments, these dosings may be reduced or increased based upon thebiological factors of a particular patient such as increased ordecreased metabolic breakdown of the drug or decreased uptake by thedigestive tract if administered orally. Additionally, the newderivatives of viridicatumtoxin may be more efficacious and thus asmaller dose is required to achieve a similar effect. Such a dose istypically administered once a day for a few weeks or until sufficientreducing in cancer cells has been achieved.

The therapeutic methods of the invention (which include prophylactictreatment) in general include administration of a therapeuticallyeffective amount of the compositions described herein to a subject inneed thereof, including a mammal, particularly a human. Such treatmentwill be suitably administered to subjects, particularly humans,suffering from, having, susceptible to, or at risk for a disease,disorder, or symptom thereof. Determination of those subjects “at risk”can be made by any objective or subjective determination by a diagnostictest or opinion of a subject or health care provider (e.g., genetictest, enzyme or protein marker, marker (as defined herein), familyhistory, and the like).

In one embodiment, the invention provides a method of monitoringtreatment progress. The method includes the step of determining a levelof changes in hematological parameters and/or cancer stem cell (CSC)analysis with cell surface proteins as diagnostic markers (which caninclude, for example, but are not limited to CD34, CD38, CD90, andCD117) or diagnostic measurement (e.g., screen, assay) in a subjectsuffering from or susceptible to a disorder or symptoms thereofassociated with cancer (e.g., leukemia) in which the subject has beenadministered a therapeutic amount of a composition as described herein.The level of marker determined in the method can be compared to knownlevels of marker in either healthy normal controls or in other afflictedpatients to establish the subject's disease status. In preferredembodiments, a second level of marker in the subject is determined at atime point later than the determination of the first level, and the twolevels are compared to monitor the course of disease or the efficacy ofthe therapy. In certain preferred embodiments, a pre-treatment level ofmarker in the subject is determined prior to beginning treatmentaccording to the methods described herein; this pre-treatment level ofmarker can then be compared to the level of marker in the subject afterthe treatment commences, to determine the efficacy of the treatment.

C. Combination Therapies

It is envisioned that the viridicatumtoxin derivatives described hereinmay be used in combination therapies with an additional antimicrobialagent such as anti-viral, antibiotic, or a compound which mitigates oneor more of the side effects experienced by the patient.

Furthermore, it is very common in the field of cancer therapy to combinetherapeutic modalities. The following is a general discussion oftherapies that may be used in conjunction with the therapies of thepresent disclosure.

To treat cancers using the methods and compositions of the presentdisclosure, one would generally contact a tumor cell or subject with acompound and at least one other therapy. These therapies would beprovided in a combined amount effective to achieve a reduction in one ormore disease parameter. This process may involve contacting thecells/subjects with the both agents/therapies at the same time, e.g.,using a single composition or pharmacological formulation that includesboth agents, or by contacting the cell/subject with two distinctcompositions or formulations, at the same time, wherein one compositionincludes the compound and the other includes the other agent.

Alternatively, viridicatumtoxin derivatives of the present disclosuremay precede or follow the other treatment by intervals ranging fromminutes to weeks. One would generally ensure that a significant periodof time did not expire between the time of each delivery, such that thetherapies would still be able to exert an advantageously combined effecton the cell/subject. In such instances, it is contemplated that onewould contact the cell with both modalities within about 12-24 hours ofeach other, within about 6-12 hours of each other, or with a delay timeof only about 12 hours. In some situations, it may be desirable toextend the time period for treatment significantly; however, whereseveral days (2, 3, 4, 5, 6 or 7) to several weeks (1, 2, 3, 4, 5, 6, 7or 8) lapse between the respective administrations.

It also is conceivable that more than one administration of either thecompound or the other therapy will be desired. Various combinations maybe employed, where a compound of the present disclosure is “A,” and theother therapy is “B,” as exemplified below:

A/B/A B/A/B B/B/A A/A/B B/A/A A/B/B B/B/B/A B/B/A/B A/A/B/B A/B/A/BA/B/B/A B/B/A/A B/A/B/A B/A/A/B B/B/B/A A/A/A/B B/A/A/A A/B/A/A A/A/B/AA/B/B/B B/A/B/B B/B/A/B

Agents or factors suitable for use in a combined therapy with agentsaccording to the present disclosure against an infectious diseaseinclude antibiotics such as penicillins, cephalosporins, carbapenems,macrolides, aminoglycosides, quinolones (including fluoroquinolones),sulfonamides and tetracylcines. In particular, one may use combinationtherapies for treating MRSA. Both CA-MRSA and HA-MRSA are resistant totraditional anti-staphylococcal beta-lactam antibiotics, such ascephalexin. CA-MRSA has a greater spectrum of antimicrobialsusceptibility, including to sulfa drugs (likeco-trimoxazole/trimethoprim-sulfamethoxazole), tetracyclines (likedoxycycline and minocycline) and clindamycin (for osteomyelitis), butthe drug of choice for treating CA-MRSA is now believed to bevancomycin, according to a Henry Ford Hospital Study. HA-MRSA isresistant even to these antibiotics and often is susceptible only tovancomycin. Newer drugs, such as linezolid (belonging to the neweroxazolidinones class) and daptomycin, are effective against both CA-MRSAand HA-MRSA. The Infectious Disease Society of America recommendsvancomycin, linezolid, or clindamycin (if susceptible) for treatingpatients with MRSA pneumonia. Ceftaroline, a fifth generationcephalosporin, is the first beta-lactam antibiotic approved in the U.S.to treat MRSA infections (skin and soft tissue or community acquiredpneumonia only).

Quinolone-resistant S. aureus is another emerging pathogen that may betreated with compounds according to the present invention, optionally incombination with vancomycin, teicoplanin and linezolid.

Vancomycin and teicoplanin are glycopeptide antibiotics used to treatMRSA infections. Teicoplanin is a structural congener of vancomycin thathas a similar activity spectrum but a longer half-life. Because the oralabsorption of vancomycin and teicoplanin is very low, these agents mustbe administered intravenously to control systemic infections. Treatmentof MRSA infection with vancomycin can be complicated, due to itsinconvenient route of administration. Moreover, many clinicians believethat the efficacy of vancomycin against MRSA is inferior to that ofanti-staphylococcal beta-lactam antibiotics againstmethicillin-susceptible Staphylococcus aureus (MSSA).

Several newly discovered strains of MRSA show antibiotic resistance evento vancomycin and teicoplanin. These new evolutions of the MRSAbacterium have been dubbed Vancomycin intermediate-resistantStaphylococcus aureus (VISA). Linezolid, quinupristin/dalfopristin,daptomycin, ceftaroline, and tigecycline are used to treat more severeinfections that do not respond to glycopeptides such as vancomycin.Current guidelines recommend daptomycin for VISA bloodstream infectionsand endocarditis.

Studies suggest that allicin, a compound found in garlic, may prove tobe effective in the treatment of MRSA.

Other combinations are contemplated. The following is a generaldiscussion of antibiotic, antiviral, and cancer therapies that may beused in combination with the compounds of the present disclosure.

1. Antibiotics

The term “antibiotics” are drugs which may be used to treat a bacterialinfection through either inhibiting the growth of bacteria or killingbacteria. Without being bound by theory, it is believed that antibioticscan be classified into two major classes: bactericidal agents that killbacteria or bacteriostatic agents that slow down or prevent the growthof bacteria.

The first commerically available antibiotic was released in the 1930's.Since then, many different antibiotics have been developed and widelyprescribed. In 2010, on average, 4 in 5 Americans are prescribedantibiotics annually. Given the prevalence of anitbiotics, bacteria havestarted to develop resistance to specific antibiotics and antibioticmechanisms. Without being bound by theory, the use of antibiotics incombination with another antibiotic may modulate resistance and enhancethe efficacy of one or both agents.

In some embodiments, antibiotics can fall into a wide range of classes.In some embodiments, the compounds of the present disclosure may be usedin conjunction with another antibiotic. In some embodiments, thecompounds may be used in conjunction with a narrow spectrum antibioticwhich targets a specific bacteria type. In some non-limiting examples ofbactericidal antibiotics include penicillin, cephalosporin, polymyxin,rifamycin, lipiarmycin, quinolones, and sulfonamides. In somenon-limiting examples of bacteriostatic antibiotics include macrolides,lincosamides, or tetracyclines. In some embodiments, the antibiotic isan aminoglycoside such as kanamycin and streptomycin, an ansamycin suchas rifaximin and geldanamycin, a carbacephem such as loracarbef, acarbapenem such as ertapenem, imipenem, a cephalosporin such ascephalexin, cefixime, cefepime, and ceftobiprole, a glycopeptide such asvancomycin or teicoplanin, a lincosamide such as lincomycin andclindamycin, a lipopeptide such as daptomycin, a macrolide such asclarithromycin, spiramycin, azithromycin, and telithromycin, amonobactam such as aztreonam, a nitrofuran such as furazolidone andnitrofurantoin, an oxazolidonones such as linezolid, a penicillin suchas amoxicillin, azlocillin, flucloxacillin, and penicillin G, anantibiotic polypeptide such as bacitracin, polymyxin B, and colistin, aquinolone such as ciprofloxacin, levofloxacin, and gatifloxacin, asulfonamide such as silver sulfadiazine, mefenide, sulfadimethoxine, orsulfasalazine, or a tetracycline such as demeclocycline, doxycycline,minocycline, oxytetracycline, or tetracycline. In some embodiments, thecompounds could be combined with a drug which acts against mycobacteriasuch as cycloserine, capreomycin, ethionamide, rifampicin, rifabutin,rifapentine, and streptomycin. Other antibiotics that are contemplatedfor combination therapies may include arsphenamine, chloramphenicol,fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin,quinupristin, dalfopristin, thiamphenicol, tigecycline, tinidazole, ortrimethoprim.

2. Antivirals

The term “antiviral” or “antiviral agents” are drugs which may be usedto treat a viral infection. In general, antiviral agents act via twomajor mechanisms: preventing viral entry into the cell and inhibitingviral synthesis. Without being bound by theory, viral replication can beinhibited by using agents that mimic either the virus-associatedproteins and thus block the cellular receptors or using agents thatmimic the cellular receptors and thus block the virus-associatedproteins. Furthermore, agents which cause an uncoating of the virus canalso be used as antiviral agents.

The second mechanism of viral inhibition is preventing or interruptingviral synthesis. Such drugs can target different proteins associatedwith the replication of viral DNA including reverse transcriptase,integrase, transcription factors, or ribozymes. Additionally, thetherapeutic agent interrupts translation by acting as an antisense DNAstrain, inhibiting the formation of protein processing or assembly, oracting as virus protease inhibitors. Finally, an anti-viral agent couldadditionally inhibit the release of the virus after viral production inthe cell.

Additionally, anti-viral agents could modulate the bodies own immunesystem to fight a viral infection. Without being bound by theory, theanti-viral agent which stimulates the immune system may be used with awide variety of viral infections.

In some embodiments, the present disclosure provides methods of usingthe disclosed compounds in a combination therapy with an anti-viralagent as described above. In some non-limiting examples, the anti-viralagent is abacavir, aciclovir, acyclovir, adefovir, amantadine,amprenavir, ampligen, arbidol, atazanavir, atripla, balavir,boceprevirertet, cidofovir, combivir, dolutegravir, daruavir,delavirdine, didanosine, docosanol, edoxudine, efavirenz, emtricitabine,enfuvirtide, entecavir, ecoliever, famciclovir, fomivirsen,fosamprenavir, foscarnet, fosfonet, ganciclovir, ibacitabine, imunovir,idoxuridine, imiquimod, indinavir, inosine, interferon type I, type II,and type III, lamivudine, lopinavir, loviride, maraviroc, moroxydine,methisazone, nelfinavir, nevirapine, nexavir, oseltamivir, penciclovir,peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin,rimantadine, ritonavir, pyramidine, saquinavir, sofosbuvir, stavudine,telaprevir, tenofovir, tenofovir disoproxil, tipranavir, trifluridine,trizivir, tromantadine, truvada, traporved, valaciclovir,valganciclovir, vicriviroc, vidarabine, viramidine, zalcitabine,zanamivir, or zidovudine. In some embodiments, the anti-viral agents isan anti-retroviral, a fusion inhibitor, an integrase inhibitor, aninterferon, a nucleoside analogues, a protease inhibitor, a reversetranscriptase inhibitor, a synergistic enhancer, or a natural productsuch as tea tree oil.

3. Chemotherapy

The term “chemotherapy” refers to the use of drugs to treat cancer. A“chemotherapeutic agent” is used to connote a compound or compositionthat is administered in the treatment of cancer. These agents or drugsare categorized by their mode of activity within a cell, for example,whether and at what stage they affect the cell cycle. Alternatively, anagent may be characterized based on its ability to directly cross-linkDNA, to intercalate into DNA, or to induce chromosomal and mitoticaberrations by affecting nucleic acid synthesis. Most chemotherapeuticagents fall into the following categories: alkylating agents,antimetabolites, antitumor antibiotics, mitotic inhibitors, andnitrosoureas.

Examples of chemotherapeutic agents include alkylating agents such asthiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan,improsulfan and piposulfan; aziridines such as benzodopa, carboquone,meturedopa, and uredopa; ethylenimines and methylamelamines includingaltretamine, triethylenemelamine, trietylenephosphoramide,triethiylenethiophosphoramide and trimethylolomelamine; acetogenins(especially bullatacin and bullatacinone); a camptothecin (including thesynthetic analogue topotecan); bryostatin; callystatin; CC-1065(including its adozelesin, carzelesin and bizelesin syntheticanalogues); cryptophycins (particularly cryptophycin 1 and cryptophycin8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin;spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine,cholophosphamide, estramustine, ifosfamide, mechlorethamine,mechlorethamine oxide hydrochloride, melphalan, novembichin,phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureassuch as carmustine, chlorozotocin, fotemustine, lomustine, nimustine,and ranimnustine; antibiotics such as the enediyne antibiotics (e.g.,calicheamicin, especially calicheamicin gammalI and calicheamicinomegaI1; dynemicin, including dynemicin A uncialamycin and derivativesthereof; bisphosphonates, such as clodronate; an esperamicin; as well asneocarzinostatin chromophore and related chromoprotein enediyneantiobiotic chromophores, aclacinomysins, actinomycin, authrarnycin,azaserine, bleomycins, cactinomycin, carabicin, carminomycin,carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin (includingmorpholino-doxorubicin, cyanomorpholino-doxorubicin,2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolicacid, nogalarnycin, olivomycins, peplomycin, potfiromycin, puromycin,quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexateand 5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adrenals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as frolinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil;amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids suchas maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol;nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK polysaccharidecomplex); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonicacid; triaziquone; 2,2′,2″-trichlorotriethylamine; trichothecenes(especially T-2 toxin, verracurin A, roridin A and anguidine); urethan;vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol;pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide;thiotepa; taxoids, e.g., paclitaxel and doxetaxel; chlorambucil;gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinumcoordination complexes such as cisplatin, oxaliplatin and carboplatin;vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone;vincristine; vinorelbine; novantrone; teniposide; edatrexate;daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11);topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO);retinoids such as retinoic acid; capecitabine; cisplatin (CDDP),carboplatin, procarbazine, mechlorethamine, cyclophosphamide,camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea,dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin,mitomycin, etoposide (VP16), tamoxifen, raloxifene, estrogen receptorbinding agents, taxol, paclitaxel, docetaxel, gemcitabien, navelbine,farnesyl-protein tansferase inhibitors, transplatinum, 5-fluorouracil,vincristin, vinblastin and methotrexate and pharmaceutically acceptablesalts, acids or derivatives of any of the above.

4. Radiotherapy

Radiotherapy, also called radiation therapy, is the treatment of cancerand other diseases with ionizing radiation. Ionizing radiation depositsenergy that injures or destroys cells in the area being treated bydamaging their genetic material, making it impossible for these cells tocontinue to grow. Although radiation damages both cancer cells andnormal cells, the latter are able to repair themselves and functionproperly.

Radiation therapy used according to the present invention may include,but is not limited to, the use of γ-rays, X-rays, and/or the directeddelivery of radioisotopes to tumor cells. Other forms of DNA damagingfactors are also contemplated such as microwaves and UV-irradiation. Itis most likely that all of these factors induce a broad range of damageon DNA, on the precursors of DNA, on the replication and repair of DNA,and on the assembly and maintenance of chromosomes. Dosage ranges forX-rays range from daily doses of 50 to 200 roentgens for prolongedperiods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens.Dosage ranges for radioisotopes vary widely, and depend on the half-lifeof the isotope, the strength and type of radiation emitted, and theuptake by the neoplastic cells.

Radiotherapy may comprise the use of radiolabeled antibodies to deliverdoses of radiation directly to the cancer site (radioimmunotherapy).Antibodies are highly specific proteins that are made by the body inresponse to the presence of antigens (substances recognized as foreignby the immune system). Some tumor cells contain specific antigens thattrigger the production of tumor-specific antibodies. Large quantities ofthese antibodies can be made in the laboratory and attached toradioactive substances (a process known as radiolabeling). Once injectedinto the body, the antibodies actively seek out the cancer cells, whichare destroyed by the cell-killing (cytotoxic) action of the radiation.This approach can minimize the risk of radiation damage to healthycells.

Conformal radiotherapy uses the same radiotherapy machine, a linearaccelerator, as the normal radiotherapy treatment but metal blocks areplaced in the path of the x-ray beam to alter its shape to match that ofthe cancer. This ensures that a higher radiation dose is given to thetumor. Healthy surrounding cells and nearby structures receive a lowerdose of radiation, so the possibility of side effects is reduced. Adevice called a multi-leaf collimator has been developed and may be usedas an alternative to the metal blocks. The multi-leaf collimatorconsists of a number of metal sheets which are fixed to the linearaccelerator. Each layer can be adjusted so that the radiotherapy beamscan be shaped to the treatment area without the need for metal blocks.Precise positioning of the radiotherapy machine is very important forconformal radiotherapy treatment and a special scanning machine may beused to check the position of internal organs at the beginning of eachtreatment.

High-resolution intensity modulated radiotherapy also uses a multi-leafcollimator. During this treatment the layers of the multi-leafcollimator are moved while the treatment is being given. This method islikely to achieve even more precise shaping of the treatment beams andallows the dose of radiotherapy to be constant over the whole treatmentarea.

Although research studies have shown that conformal radiotherapy andintensity modulated radiotherapy may reduce the side effects ofradiotherapy treatment, it is possible that shaping the treatment areaso precisely could stop microscopic cancer cells just outside thetreatment area being destroyed. This means that the risk of the cancercoming back in the future may be higher with these specializedradiotherapy techniques.

Scientists also are looking for ways to increase the effectiveness ofradiation therapy. Two types of investigational drugs are being studiedfor their effect on cells undergoing radiation. Radiosensitizers makethe tumor cells more likely to be damaged, and radioprotectors protectnormal tissues from the effects of radiation. Hyperthermia, the use ofheat, is also being studied for its effectiveness in sensitizing tissueto radiation.

5. Immunotherapy

In the context of cancer treatment, immunotherapeutics, generally, relyon the use of immune effector cells and molecules to target and destroycancer cells. Trastuzumab (Herceptin™) is such an example. The immuneeffector may be, for example, an antibody specific for some marker onthe surface of a tumor cell. The antibody alone may serve as an effectorof therapy or it may recruit other cells to actually affect cellkilling. The antibody also may be conjugated to a drug or toxin(chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussistoxin, etc.) and serve merely as a targeting agent. Alternatively, theeffector may be a lymphocyte carrying a surface molecule that interacts,either directly or indirectly, with a tumor cell target. Variouseffector cells include cytotoxic T cells and NK cells. The combinationof therapeutic modalities, i.e., direct cytotoxic activity andinhibition or reduction of ErbB2 would provide therapeutic benefit inthe treatment of ErbB2 overexpressing cancers.

In one aspect of immunotherapy, the tumor cell must bear some markerthat is amenable to targeting, i.e., is not present on the majority ofother cells. Many tumor markers exist and any of these may be suitablefor targeting in the context of the present invention. Common tumormarkers include carcinoembryonic antigen, prostate specific antigen,urinary tumor associated antigen, fetal antigen, tyrosinase (p97), gp68,TAG-72, HMFG, Sialyl Lewis Antigen, MucA, MucB, PLAP, estrogen receptor,laminin receptor, erb B and p155. An alternative aspect of immunotherapyis to combine anticancer effects with immune stimulatory effects. Immunestimulating molecules also exist including: cytokines such as IL-2,IL-4, IL-12, GM-CSF, γ-IFN, chemokines such as MIP-1, MCP-1, IL-8 andgrowth factors such as FLT3 ligand. Combining immune stimulatingmolecules, either as proteins or using gene delivery in combination witha tumor suppressor has been shown to enhance anti-tumor effects (Ju etal., 2000). Moreover, antibodies against any of these compounds may beused to target the anti-cancer agents discussed herein.

Examples of immunotherapies currently under investigation or in use areimmune adjuvants e.g., Mycobacterium bovis, Plasmodium falciparum,dinitrochlorobenzene and aromatic compounds (U.S. Pat. Nos. 5,801,005and 5,739,169; Hui and Hashimoto, 1998; Christodoulides et al., 1998),cytokine therapy, e.g., interferons α, β, and γ; IL-1, GM-CSF and TNF(Bukowski et al., 1998; Davidson et al., 1998; Hellstrand et al., 1998)gene therapy, e.g., TNF, IL-1, IL-2, p53 (Qin et al., 1998; Austin-Wardand Villaseca, 1998; U.S. Pat. Nos. 5,830,880 and 5,846,945) andmonoclonal antibodies, e.g., anti-ganglioside GM2, anti-HER-2, anti-p185(Pietras et al., 1998; Hanibuchi et al., 1998; U.S. Pat. No. 5,824,311).It is contemplated that one or more anti-cancer therapies may beemployed with the gene silencing therapies described herein.

In active immunotherapy, an antigenic peptide, polypeptide or protein,or an autologous or allogenic tumor cell composition or “vaccine” isadministered, generally with a distinct bacterial adjuvant (Ravindranathand Morton, 1991; Morton et al., 1992; Mitchell et al., 1990; Mitchellet al., 1993).

In adoptive immunotherapy, the patient's circulating lymphocytes, ortumor infiltrated lymphocytes, are isolated in vitro, activated bylymphokines such as IL-2 or transduced with genes for tumor necrosis,and readministered (Rosenberg et al., 1988; 1989).

6. Surgery

Approximately 60% of persons with cancer will undergo surgery of sometype, which includes preventative, diagnostic or staging, curative, andpalliative surgery. Curative surgery is a cancer treatment that may beused in conjunction with other therapies, such as the treatment of thepresent invention, chemotherapy, radiotherapy, hormonal therapy, genetherapy, immunotherapy and/or alternative therapies.

Curative surgery includes resection in which all or part of canceroustissue is physically removed, excised, and/or destroyed. Tumor resectionrefers to physical removal of at least part of a tumor. In addition totumor resection, treatment by surgery includes laser surgery,cryosurgery, electrosurgery, and microscopically controlled surgery(Mohs' surgery). It is further contemplated that the present inventionmay be used in conjunction with removal of superficial cancers,precancers, or incidental amounts of normal tissue.

Upon excision of part or all of cancerous cells, tissue, or tumor, acavity may be formed in the body. Treatment may be accomplished byperfusion, direct injection or local application of the area with anadditional anti-cancer therapy. Such treatment may be repeated, forexample, every 1, 2, 3, 4, 5, 6, or 7 days, or every 1, 2, 3, 4, and 5weeks or every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. Thesetreatments may be of varying dosages as well.

In some particular embodiments, after removal of the tumor, an adjuvanttreatment with a compound of the present disclosure is believe to beparticularly efficacious in reducing the reoccurance of the tumor.Additionally, the compounds of the present disclosure can also be usedin a neoadjuvant setting.

7. Other Agents

It is contemplated that other agents may be used with the presentinvention. These additional agents include immunomodulatory agents,agents that affect the upregulation of cell surface receptors and GAPjunctions, cytostatic and differentiation agents, inhibitors of celladhesion, agents that increase the sensitivity of the hyperproliferativecells to apoptotic inducers, or other biological agents.Immunomodulatory agents include tumor necrosis factor; interferon alpha,beta, and gamma; IL-2 and other cytokines; F42K and other cytokineanalogs; or MIP-1, MIP-1β, MCP-1, RANTES, and other chemokines. It isfurther contemplated that the upregulation of cell surface receptors ortheir ligands such as Fas/Fas ligand, DR4 or DR5/TRAIL (Apo-2 ligand)would potentiate the apoptotic inducing abilities of the presentinvention by establishment of an autocrine or paracrine effect onhyperproliferative cells. Increases intercellular signaling by elevatingthe number of GAP junctions would increase the anti-hyperproliferativeeffects on the neighboring hyperproliferative cell population. In otherembodiments, cytostatic or differentiation agents may be used incombination with the present invention to improve theanti-hyerproliferative efficacy of the treatments. Inhibitors of celladhesion are contemplated to improve the efficacy of the presentinvention. Examples of cell adhesion inhibitors are focal adhesionkinase (FAKs) inhibitors and Lovastatin. It is further contemplated thatother agents that increase the sensitivity of a hyperproliferative cellto apoptosis, such as the antibody c225, could be used in combinationwith the present invention to improve the treatment efficacy.

There have been many advances in the therapy of cancer following theintroduction of cytotoxic chemotherapeutic drugs. However, one of theconsequences of chemotherapy is the development/acquisition ofdrug-resistant phenotypes and the development of multiple drugresistance. The development of drug resistance remains a major obstaclein the treatment of such tumors and therefore, there is an obvious needfor alternative approaches such as gene therapy.

Another form of therapy for use in conjunction with chemotherapy,radiation therapy or biological therapy includes hyperthermia, which isa procedure in which a patient's tissue is exposed to high temperatures(up to 106° F.). External or internal heating devices may be involved inthe application of local, regional, or whole-body hyperthermia. Localhyperthermia involves the application of heat to a small area, such as atumor. Heat may be generated externally with high-frequency wavestargeting a tumor from a device outside the body. Internal heat mayinvolve a sterile probe, including thin, heated wires or hollow tubesfilled with warm water, implanted microwave antennae, or radiofrequencyelectrodes.

A patient's organ or a limb is heated for regional therapy, which isaccomplished using devices that produce high energy, such as magnets.Alternatively, some of the patient's blood may be removed and heatedbefore being perfused into an area that will be internally heated.Whole-body heating may also be implemented in cases where cancer hasspread throughout the body. Warm-water blankets, hot wax, inductivecoils, and thermal chambers may be used for this purpose.

The skilled artisan is directed to “Remington's Pharmaceutical Sciences”15th Edition, chapter 33, in particular pages 624-652. Some variation indosage will necessarily occur depending on the condition of the subjectbeing treated. The person responsible for administration will, in anyevent, determine the appropriate dose for the individual subject.Moreover, for human administration, preparations should meet sterility,pyrogenicity, general safety and purity standards as required by FDAOffice of Biologics standards.

It also should be pointed out that any of the foregoing therapies mayprove useful by themselves in treating cancer.

IV. SYNTHETIC METHODS

In some aspects, the compounds of this invention can be synthesizedusing the methods of organic chemistry as described in this application.These methods can be further modified and optimized using the principlesand techniques of organic chemistry as applied by a person skilled inthe art. Such principles and techniques are taught, for example, inMarch's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure(2007), which is incorporated by reference herein

A. Process Scale-Up

The synthetic methods described herein can be further modified andoptimized for preparative, pilot- or large-scale production, eitherbatch of continuous, using the principles and techniques of processchemistry as applied by a person skilled in the art. Such principles andtechniques are taught, for example, in Practical Process Research &Development (2000), which is incorporated by reference herein. Thesynthetic method described herein may be used to produce preparativescale amounts of viridicatumtoxin and derivatives thereof.

B. Chemical Definitions

When used in the context of a chemical group: “hydrogen” means —H;“hydroxy” means —OH; “oxo” means ═O; “carbonyl” means —C(═O)—; “carboxy”means —C(═O)OH (also written as —COOH or —CO₂H); “halo” meansindependently —F, —Cl, —Br or —I; “amino” means —NH₂; “hydroxyamino”means —NHOH; “nitro” means —NO₂; imino means ═NH; “cyano” means —CN;“isocyanate” means —N═C═O; “azido” means —N₃; in a monovalent context“phosphate” means —OP(O)(OH)₂ or a deprotonated form thereof; in adivalent context “phosphate” means —OP(O)(OH)O— or a deprotonated formthereof; “mercapto” means —SH; and “thio” means ═S; “sulfo” means —SO₃H,“sulfonyl” means —S(O)₂—; and “sulfinyl” means —S(O)—.

In the context of chemical formulas, the symbol “—” means a single bond,“═” means a double bond, and “≡” means triple bond. The symbol “

” represents an optional bond, which if present is either single ordouble. The symbol “

” represents a single bond or a double bond. Thus, for example, theformula

includes

And it is understood that no one such ring atom forms part of more thanone double bond. Furthermore, it is noted that the covalent bond symbol“—”, when connecting one or two stereogenic atoms, does not indicate anypreferred stereochemistry. Instead, it covers all stereoisomers as wellas mixtures thereof. The symbol “

”, when drawn perpendicularly across a bond

for methyl) indicates a point of attachment of the group. It is notedthat the point of attachment is typically only identified in this mannerfor larger groups in order to assist the reader in unambiguouslyidentifying a point of attachment. The symbol “

” means a single bond where the group attached to the thick end of thewedge is “out of the page.” The symbol “

” means a single bond where the group attached to the thick end of thewedge is “into the page”. The symbol “

” means a single bond where the geometry around a double bond (e.g.,either E or Z) is undefined. Both options, as well as combinationsthereof are therefore intended. Any undefined valency on an atom of astructure shown in this application implicitly represents a hydrogenatom bonded to that atom. A bold dot on a carbon atom indicates that thehydrogen attached to that carbon is oriented out of the plane of thepaper.

When a group “R” is depicted as a “floating group” on a ring system, forexample, in the formula:

then R may replace any hydrogen atom attached to any of the ring atoms,including a depicted, implied, or expressly defined hydrogen, so long asa stable structure is formed. When a group “R” is depicted as a“floating group” on a fused ring system, as for example in the formula:

then R may replace any hydrogen attached to any of the ring atoms ofeither of the fused rings unless specified otherwise. Replaceablehydrogens include depicted hydrogens (e.g., the hydrogen attached to thenitrogen in the formula above), implied hydrogens (e.g., a hydrogen ofthe formula above that is not shown but understood to be present),expressly defined hydrogens, and optional hydrogens whose presencedepends on the identity of a ring atom (e.g., a hydrogen attached togroup X, when X equals —CH—), so long as a stable structure is formed.In the example depicted, R may reside on either the 5-membered or the6-membered ring of the fused ring system. In the formula above, thesubscript letter “y” immediately following the group “R” enclosed inparentheses, represents a numeric variable. Unless specified otherwise,this variable can be 0, 1, 2, or any integer greater than 2, onlylimited by the maximum number of replaceable hydrogen atoms of the ringor ring system.

For the groups and classes below, the following parenthetical subscriptsfurther define the group/class as follows: “(Cn)” defines the exactnumber (n) of carbon atoms in the group/class. “(C≦n)” defines themaximum number (n) of carbon atoms that can be in the group/class, withthe minimum number as small as possible for the group in question, e.g.,it is understood that the minimum number of carbon atoms in the group“alkenyl_((C≦8))” or the class “alkene_((C≦8))” is two. For example,“alkoxy_((C≦10))” designates those alkoxy groups having from 1 to 10carbon atoms. (Cn−n′) defines both the minimum (n) and maximum number(n′) of carbon atoms in the group. Similarly, “alkyl_((C2-10))”designates those alkyl groups having from 2 to 10 carbon atoms.

The term “saturated” as used herein means the compound or group somodified has no carbon-carbon double and no carbon-carbon triple bonds,except as noted below. In the case of substituted versions of saturatedgroups, one or more carbon oxygen double bond or a carbon nitrogendouble bond may be present. And when such a bond is present, thencarbon-carbon double bonds that may occur as part of keto-enoltautomerism or imine/enamine tautomerism are not precluded.

The term “aliphatic” when used without the “substituted” modifiersignifies that the compound/group so modified is an acyclic or cyclic,but non-aromatic hydrocarbon compound or group. In aliphaticcompounds/groups, the carbon atoms can be joined together in straightchains, branched chains, or non-aromatic rings (alicyclic). Aliphaticcompounds/groups can be saturated, that is joined by single bonds(alkanes/alkyl), or unsaturated, with one or more double bonds(alkenes/alkenyl) or with one or more triple bonds (alkynes/alkynyl).

The term “alkyl” when used without the “substituted” modifier refers toa monovalent saturated aliphatic group with a carbon atom as the pointof attachment, a linear or branched acyclic structure, and no atomsother than carbon and hydrogen. The groups —CH₃ (Me), —CH₂CH₃ (Et),—CH₂CH₂CH₃ (n-Pr or propyl), —CH(CH₃)₂ (i-Pr, ^(i)Pr or isopropyl),—CH₂CH₂CH₂CH₃ (n-Bu), —CH(CH₃)CH₂CH₃ (sec-butyl), —CH₂CH(CH₃)₂(isobutyl), —C(CH₃)₃ (tert-butyl, t-butyl, t-Bu or ^(t)Bu), and—CH₂C(CH₃)₃ (neo-pentyl) are non-limiting examples of alkyl groups. Theterm “alkanediyl” when used without the “substituted” modifier refers toa divalent saturated aliphatic group, with one or two saturated carbonatom(s) as the point(s) of attachment, a linear or branched acyclicstructure, no carbon-carbon double or triple bonds, and no atoms otherthan carbon and hydrogen. The groups, —CH₂-(methylene), —CH₂CH₂—,—CH₂C(CH₃)₂CH₂—, and —CH₂CH₂CH₂—, are non-limiting examples ofalkanediyl groups. The term “alkylidene” when used without the“substituted” modifier refers to the divalent group ═CRR′ in which R andR′ are independently hydrogen or alkyl. Non-limiting examples ofalkylidene groups include: ═CH₂, ═CH(CH₂CH₃), and ═C(CH₃)₂. An “alkane”refers to the compound H—R, wherein R is alkyl as this term is definedabove. When any of these terms is used with the “substituted” modifierone or more hydrogen atom has been independently replaced by —OH, —F,—Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃,—C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. The following groups are non-limiting examples of substitutedalkyl groups: —CH₂OH, —CH₂Cl, —CF₃, —CH₂CN, —CH₂C(O)OH, —CH₂C(O)OCH₃,—CH₂C(O)NH₂, —CH₂C(O)CH₃, —CH₂OCH₃, —CH₂OC(O)CH₃, —CH₂NH₂, —CH₂N(CH₃)₂,and —CH₂CH₂Cl. The term “haloalkyl” is a subset of substituted alkyl, inwhich one or more hydrogen atoms has been substituted with a halo groupand no other atoms aside from carbon, hydrogen and halogen are present.The group, —CH₂Cl is a non-limiting example of a haloalkyl. The term“fluoroalkyl” is a subset of substituted alkyl, in which one or morehydrogen has been substituted with a fluoro group and no other atomsaside from carbon, hydrogen and fluorine are present. The groups, —CHF,—CF₃, and —CH₂CF₃ are non-limiting examples of fluoroalkyl groups.

The term “cycloalkyl” when used without the “substituted” modifierrefers to a monovalent saturated aliphatic group with a carbon atom asthe point of attachment, said carbon atom forms part of one or morenon-aromatic ring structures, a cyclo or cyclic structure, nocarbon-carbon double or triple bonds, and no atoms other than carbon andhydrogen. Non-limiting examples of cycloalkyl groups include: —CH(CH₂)₂(cyclopropyl), cyclobutyl, cyclopentyl, or cyclohexyl. The term“cycloalkanediyl” when used without the “substituted” modifier refers toa divalent saturated aliphatic group with one or two carbon atom as thepoint(s) of attachment, said carbon atom(s) forms part of one or morenon-aromatic ring structures, a cyclo or cyclic structure, nocarbon-carbon double or triple bonds, and no atoms other than carbon andhydrogen.

are non-limiting examples of cycloalkanediyl groups. A “cycloalkane”refers to the compound H—R, wherein R is cycloalkyl as this term isdefined above. When any of these terms is used with the “substituted”modifier one or more hydrogen atom has been independently replaced by—OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃,—OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. The following groups are non-limiting examples of substitutedcycloalkyl groups: —C(OH)(CH₂)₂,

The term “alkenyl” when used without the “substituted” modifier refersto a monovalent unsaturated aliphatic group with a carbon atom as thepoint of attachment, a linear or branched, acyclic structure, at leastone nonaromatic carbon-carbon double bond, no carbon-carbon triplebonds, and no atoms other than carbon and hydrogen. Non-limitingexamples of alkenyl groups include: —CH═CH₂ (vinyl), —CH═CHCH₃,—CH═CHCH₂CH₃, —CH₂CH═CH₂ (allyl), —CH₂CH═CHCH₃, and —CH═CHCH═CH₂. Theterm “alkenediyl” when used without the “substituted” modifier refers toa divalent unsaturated aliphatic group, with two carbon atoms as pointsof attachment, a linear or branched, cyclo, cyclic or acyclic structure,at least one nonaromatic carbon-carbon double bond, no carbon-carbontriple bonds, and no atoms other than carbon and hydrogen. The groups,—CH═CH—, —CH═C(CH₃)CH₂—, and —CH═CHCH₂—, are non-limiting examples ofalkenediyl groups. It is noted that while the alkenediyl group isaliphatic, once connected at both ends, this group is not precluded fromforming part of an aromatic structure. The terms “alkene” and refer to acompound having the formula HR, wherein R is alkenyl as this term isdefined above. A “terminal alkene” refers to an alkene having just onecarbon-carbon double bond, wherein that bond forms a vinyl group at oneend of the molecule. When any of these terms are used with the“substituted” modifier one or more hydrogen atom has been independentlyreplaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH,—OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂,—OC(O)CH₃, or —S(O)₂NH₂. The groups, —CH═CHF, —CH═CHCl and —CH═CHBr, arenon-limiting examples of substituted alkenyl groups.

The term “cycloalkenyl” when used without the “substituted” modifierrefers to a monovalent unsaturated aliphatic group with a carbon atom asthe point of attachment, said carbon atom forms part of one or morenon-aromatic ring structures, a cyclo or cyclic structure, at least onenon-aromatic carbon-carbon double bond, no carbon-carbon triple bonds,and no atoms other than carbon and hydrogen. In some non-limitingexamples of cycloalkenyl groups include

The term “cycloalkenediyl” when used without the “substituted” modifierrefers to a divalent unsaturated aliphatic group with one or two carbonatom(s) as the point(s) of attachment, said carbon atom(s) forms part ofone or more non-aromatic ring structures, a cyclo or cyclic structure,at least one non-aromatic carbon-carbon double bond, no carbon-carbontriple bonds, and no atoms other than carbon and hydrogen.

are non-limiting examples of cycloalkenediyl. It is noted that while thecycloalkenediyl group is aliphatic, once connected at both ends, thisgroup is not precluded from forming part of an aromatic structure. Theterms “cycloalkene” and refer to a compound having the formula H—R,wherein R is cycloalkenyl as this term is defined above. The term“olefin” is synonymous with the terms “alkene” or a “cycloalkane” asthose terms are defined above. When any of these terms are used with the“substituted” modifier one or more hydrogen atom has been independentlyreplaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH,—OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂,—OC(O)CH₃, or —S(O)₂NH₂. In some non-limiting examples of substitutedcycloalkenyl include

The term “alkynyl” when used without the “substituted” modifier refersto a monovalent unsaturated aliphatic group with a carbon atom as thepoint of attachment, a linear or branched, acyclic structure, at leastone carbon-carbon triple bond, and no atoms other than carbon andhydrogen. As used herein, the term alkynyl does not preclude thepresence of one or more non-aromatic carbon-carbon double bonds. Thegroups, —C≡CH, —C≡CCH₃, and —CH₂C≡CCH₃, are non-limiting examples ofalkynyl groups. An “alkyne” refers to the compound H—R, wherein R isalkynyl. When any of these terms are used with the “substituted”modifier one or more hydrogen atom has been independently replaced by—OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃,—OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂.

The term “aryl” when used without the “substituted” modifier refers to amonovalent unsaturated aromatic group with an aromatic carbon atom asthe point of attachment, said carbon atom forming part of a one or moresix-membered aromatic ring structure, wherein the ring atoms are allcarbon, and wherein the group consists of no atoms other than carbon andhydrogen. If more than one ring is present, the rings may be fused orunfused. As used herein, the term does not preclude the presence of oneor more alkyl or aralkyl groups (carbon number limitation permitting)attached to the first aromatic ring or any additional aromatic ringpresent. Non-limiting examples of aryl groups include phenyl (Ph),methylphenyl, (dimethyl)phenyl, —C₆H₄CH₂CH₃ (ethylphenyl), naphthyl, anda monovalent group derived from biphenyl. The term “arenediyl” when usedwithout the “substituted” modifier refers to a divalent aromatic groupwith two aromatic carbon atoms as points of attachment, said carbonatoms forming part of one or more six-membered aromatic ringstructure(s) wherein the ring atoms are all carbon, and wherein themonovalent group consists of no atoms other than carbon and hydrogen. Asused herein, the term does not preclude the presence of one or morealkyl, aryl or aralkyl groups (carbon number limitation permitting)attached to the first aromatic ring or any additional aromatic ringpresent. If more than one ring is present, the rings may be fused orunfused. Unfused rings may be connected via one or more of thefollowing: a covalent bond, alkanediyl, or alkenediyl groups (carbonnumber limitation permitting). Non-limiting examples of arenediyl groupsinclude:

An “arene” refers to the compound HR, wherein R is aryl as that term isdefined above. Benzene and toluene are non-limiting examples of arenes.When any of these terms are used with the “substituted” modifier one ormore hydrogen atom has been independently replaced by —OH, —F, —Cl, —Br,—I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃,—NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂.

The term “aralkyl” when used without the “substituted” modifier refersto the monovalent group -alkanediyl-aryl, in which the terms alkanediyland aryl are each used in a manner consistent with the definitionsprovided above. Non-limiting examples of aralkyls are: phenylmethyl(benzyl, Bn) and 2-phenyl-ethyl. When the term aralkyl is used with the“substituted” modifier one or more hydrogen atom from the alkanediyland/or the aryl group has been independently replaced by —OH, —F, —Cl,—Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃,—C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. Non-limiting examples of substituted aralkyls are:(3-chlorophenyl)-methyl, and 2-chloro-2-phenyl-eth-1-yl.

The term “heteroaryl” when used without the “substituted” modifierrefers to a monovalent aromatic group with an aromatic carbon atom ornitrogen atom as the point of attachment, said carbon atom or nitrogenatom forming part of one or more aromatic ring structures wherein atleast one of the ring atoms is nitrogen, oxygen or sulfur, and whereinthe heteroaryl group consists of no atoms other than carbon, hydrogen,aromatic nitrogen, aromatic oxygen and aromatic sulfur. If more than onering is present, the rings may be fused or unfused. As used herein, theterm does not preclude the presence of one or more alkyl, aryl, and/oraralkyl groups (carbon number limitation permitting) attached to thearomatic ring or aromatic ring system. Non-limiting examples ofheteroaryl groups include furanyl, imidazolyl, indolyl, indazolyl (Im),isoxazolyl, methylpyridinyl, oxazolyl, phenylpyridinyl, pyridinyl,pyrrolyl, pyrimidinyl, pyrazinyl, quinolyl, quinazolyl, quinoxalinyl,triazinyl, tetrazolyl, thiazolyl, thienyl, and triazolyl. The term“N-heteroaryl” refers to a heteroaryl group with a nitrogen atom as thepoint of attachment. The term “heteroarenediyl” when used without the“substituted” modifier refers to an divalent aromatic group, with twoaromatic carbon atoms, two aromatic nitrogen atoms, or one aromaticcarbon atom and one aromatic nitrogen atom as the two points ofattachment, said atoms forming part of one or more aromatic ringstructure(s) wherein at least one of the ring atoms is nitrogen, oxygenor sulfur, and wherein the divalent group consists of no atoms otherthan carbon, hydrogen, aromatic nitrogen, aromatic oxygen and aromaticsulfur. If more than one ring is present, the rings may be fused orunfused. Unfused rings may be connected via one or more of thefollowing: a covalent bond, alkanediyl, or alkenediyl groups (carbonnumber limitation permitting). As used herein, the term does notpreclude the presence of one or more alkyl, aryl, and/or aralkyl groups(carbon number limitation permitting) attached to the aromatic ring oraromatic ring system. Non-limiting examples of heteroarenediyl groupsinclude:

A “heteroarene” refers to the compound HR, wherein R is heteroaryl.Pyridine and quinoline are non-limiting examples of heteroarenes. Whenthese terms are used with the “substituted” modifier one or morehydrogen atom has been independently replaced by —OH, —F, —Cl, —Br, —I,—NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃,—NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂.

The term “heteroaralkyl” when used without the “substituted” modifierrefers to the monovalent group -alkanediyl-heteroaryl, in which theterms alkanediyl and heteroaryl are each used in a manner consistentwith the definitions provided above. Non-limiting examples ofheteroaralkyls are: 2-pyridylmethyl and 2-indazolyl-ethyl. When the termheteroaralkyl is used with the “substituted” modifier one or morehydrogen atom from the alkanediyl and/or the heteroaryl group has beenindependently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂. Non-limiting examples ofsubstituted heteroaralkyls are: (3-chloroquinolyl)-methyl, and2-chloro-2-thienyl-eth-1-yl.

The term “heterocycloalkyl” when used without the “substituted” modifierrefers to a monovalent non-aromatic group with a carbon atom or nitrogenatom as the point of attachment, said carbon atom or nitrogen atomforming part of one or more non-aromatic ring structures wherein atleast one of the ring atoms is nitrogen, oxygen or sulfur, and whereinthe heterocycloalkyl group consists of no atoms other than carbon,hydrogen, nitrogen, oxygen and sulfur. If more than one ring is present,the rings may be fused or unfused. As used herein, the term does notpreclude the presence of one or more alkyl groups (carbon numberlimitation permitting) attached to the ring or ring system. Also, theterm does not preclude the presence of one or more double bonds in thering or ring system, provided that the resulting group remainsnon-aromatic. Non-limiting examples of heterocycloalkyl groups includeaziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydrothiofuranyl,tetrahydropyranyl, pyranyl, oxiranyl, and oxetanyl. The term“N-heterocycloalkyl” refers to a heterocycloalkyl group with a nitrogenatom as the point of attachment. The term “heterocycloalkanediyl” whenused without the “substituted” modifier refers to an divalent cyclicgroup, with two carbon atoms, two nitrogen atoms, or one carbon atom andone nitrogen atom as the two points of attachment, said atoms formingpart of one or more ring structure(s) wherein at least one of the ringatoms is nitrogen, oxygen or sulfur, and wherein the divalent groupconsists of no atoms other than carbon, hydrogen, nitrogen, oxygen andsulfur. If more than one ring is present, the rings may be fused orunfused. Unfused rings may be connected via one or more of thefollowing: a covalent bond, alkanediyl, or alkenediyl groups (carbonnumber limitation permitting). As used herein, the term does notpreclude the presence of one or more alkyl groups (carbon numberlimitation permitting) attached to the ring or ring system. Also, theterm does not preclude the presence of one or more double bonds in thering or ring system, provided that the resulting group remainsnon-aromatic. Non-limiting examples of heterocycloalkanediyl groupsinclude:

When these terms are used with the “substituted” modifier one or morehydrogen atom has been independently replaced by —OH, —F, —Cl, —Br, —I,—NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃,—NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, —S(O)₂NH₂, or —C(O)OC(CH₃)₃(tert-butyloxycarbonyl, BOC).

The term “acyl” when used without the “substituted” modifier refers tothe group —C(O)R, in which R is a hydrogen, alkyl, cycloalkyl, aryl,aralkyl or heteroaryl, as those terms are defined above. The groups,—CHO, —C(O)CH₃ (acetyl, Ac), —C(O)CH₂CH₃, —C(O)CH₂CH₂CH₃, —C(O)CH(CH₃)₂,—C(O)CH(CH₂)₂, —C(O)C₆H₅, —C(O)C₆H₄CH₃, —C(O)CH₂C₆H₅, —C(O)(imidazolyl)are non-limiting examples of acyl groups. A “thioacyl” is defined in ananalogous manner, except that the oxygen atom of the group —C(O)R hasbeen replaced with a sulfur atom, —C(S)R. The term “aldehyde”corresponds to an alkane, as defined above, wherein at least one of thehydrogen atoms has been replaced with a —CHO group. When any of theseterms are used with the “substituted” modifier one or more hydrogen atom(including a hydrogen atom directly attached the carbonyl orthiocarbonyl group, if any) has been independently replaced by —OH, —F,—Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃,—C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. The groups, —C(O)CH₂CF₃, —CO₂H (carboxyl), —CO₂CH₃(methylcarboxyl), —CO₂CH₂CH₃, —C(O)NH₂ (carbamoyl), and —CON(CH₃)₂, arenon-limiting examples of substituted acyl groups.

The term “alkylamino” when used without the “substituted” modifierrefers to the group —NHR, in which R is an alkyl, as that term isdefined above. Non-limiting examples of alkylamino groups include:—NHCH₃ and —NHCH₂CH₃. The term “dialkylamino” when used without the“substituted” modifier refers to the group —NRR′, in which R and R′ caneach independently be the same or different alkyl groups, or R and R′can be taken together to represent an alkanediyl. Non-limiting examplesof dialkylamino groups include: —N(CH₃)₂, —N(CH₃)(CH₂CH₃), andN-pyrrolidinyl. The terms “alkoxyamino”, “cycloalkylamino”,“alkenylamino”, “cycloalkenylamino”, “alkynylamino”, “arylamino”,“aralkylamino”, “heteroarylamino”, “heterocycloalkylamino” and“alkylsulfonylamino” when used without the “substituted” modifier,refers to groups, defined as —NHR, in which R is alkoxy, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, heteroaryl,heterocycloalkyl, and alkylsulfonyl, respectively. A non-limitingexample of an arylamino group is —NHC₆H₅. The term “amido” (acylamino),when used without the “substituted” modifier, refers to the group —NHR,in which R is acyl, as that term is defined above. A non-limitingexample of an amido group is —NHC(O)CH₃. The term “alkylimino” when usedwithout the “substituted” modifier refers to the divalent group ═NR, inwhich R is an alkyl, as that term is defined above. The term“alkylaminodiyl” refers to the divalent group —NH-alkanediyl-,—NH-alkanediyl-NH—, or -alkanediyl-NH-alkanediyl-. When any of theseterms is used with the “substituted” modifier one or more hydrogen atomhas been independently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂,—CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂. The groups —NHC(O)OCH₃ and—NHC(O)NHCH₃ are non-limiting examples of substituted amido groups.

The term “alkoxy” when used without the “substituted” modifier refers tothe group —OR, in which R is an alkyl, as that term is defined above.Non-limiting examples include: —OCH₃ (methoxy), —OCH₂CH₃ (ethoxy),—OCH₂CH₂CH₃, —OCH(CH₃)₂ (isopropoxy), and OC(CH₃)₃ (tert-butoxy). Theterms “cycloalkoxy”, “alkenyloxy”, “alkynyloxy”, “aryloxy”, “aralkoxy”,“heteroaryloxy”, “heterocycloalkoxy”, and “acyloxy”, when used withoutthe “substituted” modifier, refers to groups, defined as —OR, in which Ris cycloalkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,heterocycloalkyl, and acyl, respectively. The term “alkoxydiyl” refersto the divalent group —O-alkanediyl-, —O-alkanediyl-O—, or-alkanediyl-O-alkanediyl-. The term “alkylthio” and “acylthio” when usedwithout the “substituted” modifier refers to the group —SR, in which Ris an alkyl and acyl, respectively. The term “alcohol” corresponds to analkane, as defined above, wherein at least one of the hydrogen atoms hasbeen replaced with a hydroxy group. The term “ether” corresponds to analkane or cycloalkane, as defined above, wherein at least one of thehydrogen atoms has been replaced with an alkoxy or cycloalkoxy group.When any of these terms is used with the “substituted” modifier one ormore hydrogen atom has been independently replaced by —OH, —F, —Cl, —Br,—I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃,—NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂.

A “reducing agent” in the context of this application is a compoundwhich causes the reduction of a compound through the donation of anelectron. A soft reducing agent is a reducing agent which containselectron delocalizing ligands which weaken the nucleophilic strength ofthe hydride. Some non-limiting examples of reducing agents are sodiumborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride,lithium aluminum hydride, diisobutylaluminum hydride, hydrogen gas, ormetal hydride.

A “base” in the context of this application is a compound which has alone pair of electron that can accept a proton. Non-limiting examples ofa base can include triethylamine, a metal hydroxide, a metal alkoxide, ametal hydride, or a metal alkane. An alkyllithium or organolithium is acompound of the formula alkyl_((C≦12))-Li. A nitrogenous base is analkylamine, dialkylamino, trialkylamine, nitrogen containingheterocycloalkane or heteroarene wherein the base can accept a proton toform a positively charged species. For example, but not limited to, anitrogenous base could be 4,4-dimethylpyridine, pyridine,1,8-diazabicyclo[5.4.0]undec-7-ene, diisopropylethylamine, ortriethylamine. A metal alkoxide is an alkoxy group wherein rather thanthe oxygen atom which was the point of connectivity has an extraelectron and thus a negative charge which is charged balanced by themetal ion. For example, a metal alkoxide could be a sodium tert-butoxideor potassium methoxide.

An “oxidizing agent” in the context of this application is a compoundwhich causes the oxidation of a compound by accepting an electron. Somenon-limiting examples of oxidizing agent are oxygen gas, peroxides,chlorite, hypochlorite, or a chromium compound such as pyridiniumchlorochromate or hydrochromic acid.

A “metal” in the context of this application is a transition metal or ametal of groups I or II. In some embodiments, a metal is lithium,sodium, or potassium. In other embodiments, a metal is calcium ormagnesium.

An “alkylaluminium” in the context of this application is a reagentwhich contains one, two, three, or four alkyl groups as that group isdefined above to a central aluminum atom. Some non-limiting examples ofalkylaluminiums are trimethylaluminum or tetramethylaluminium.

A “linker” in the context of this application is divalent chemical groupwhich may be used to join one or more molecules to the compound of theinstant disclosure. In some embodiments, the linker contains a reactivefunctional group, such as a carboxyl, an amide, a amine, a hydroxy, amercapto, an aldehyde, or a ketone on each end that be used to join oneor more molecules to the compounds of the instant disclosure. In somenon-limiting examples, —CH₂CH₂CH₂CH₂—, —C(O)CH₂CH₂CH₂—, —OCH₂CH₂NH,—NHCH₂CH₂NH—, and —(OCH₂CH₂)_(n)— wherein n is between 1-1000, arelinkers.

A “stereoisomer” or “optical isomer” is an isomer of a given compound inwhich the same atoms are bonded to the same other atoms, but where theconfiguration of those atoms in three dimensions differs. “Enantiomers”are stereoisomers of a given compound that are mirror images of eachother, like left and right hands. “Diastereomers” are stereoisomers of agiven compound that are not enantiomers. Chiral molecules contain achiral center, also referred to as a stereocenter or stereogenic center,which is any point, though not necessarily an atom, in a moleculebearing groups such that an interchanging of any two groups leads to astereoisomer. In organic compounds, the chiral center is typically acarbon, phosphorus or sulfur atom, though it is also possible for otheratoms to be stereocenters in organic and inorganic compounds. A moleculecan have multiple stereocenters, giving it many stereoisomers. Incompounds whose stereoisomerism is due to tetrahedral stereogeniccenters (e.g., tetrahedral carbon), the total number of hypotheticallypossible stereoisomers will not exceed 2^(n), where n is the number oftetrahedral stereocenters. Molecules with symmetry frequently have fewerthan the maximum possible number of stereoisomers. A 50:50 mixture ofenantiomers is referred to as a racemic mixture. Alternatively, amixture of enantiomers can be enantiomerically enriched so that oneenantiomer is present in an amount greater than 50%. Typically,enantiomers and/or diastereomers can be resolved or separated usingtechniques known in the art. It is contemplated that that for anystereocenter or axis of chirality for which stereochemistry has not beendefined, that stereocenter or axis of chirality can be present in its Rform, S form, or as a mixture of the R and S forms, including racemicand non-racemic mixtures. As used herein, the phrase “substantially freefrom other stereoisomers” means that the composition contains ≦15%, morepreferably ≦10%, even more preferably ≦5%, or most preferably ≦1% ofanother stereoisomer(s).

V. EXAMPLES

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventor to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the spirit and scope ofthe invention.

Example 1—Synthesis of Viridicatumtoxin and Analogs Thereof

The synthesis of viridicatumtoxin analogs V2-V6 is based on the recentlydisclosed synthetic approach to racemic viridicatumtoxin B [(±)-1] asshown in scheme 1. (Nicolaou, et al., 2013, which is incorporated hereinby reference) Specifically, intermediate (±)-11 of our synthetic route(see scheme 1) was hydrogenated in the presence of palladium black toprovide access to analog (±)-V2 (scheme 2). Intermediate (±)-15-epi-11,which had been obtained as a side-product in our synthesis ofviridicatumtoxin B [(±)-1] (see scheme 1), was transformed into analog(±)-V2 accordingly (scheme 2). Intermediate (±)-15-epi-14, which wasobtained analogously to (±)-15-epi-11, (Nicolaou, et al., 2014) wassubjected to hydrogenolysis in the presence of palladium black toprovide access to analog (±)-V4 (scheme 2). For all of these analogs,ketal hydrolysis occurred during HPLC purification.

The synthesis of analogs (±)-V5 and (±)-V6 commenced with intermediate(±)-11 (scheme 3). Reduction with NaCNBH₃ (4.0 equiv.) in acetic acidled to a mixture of methyl ethers (±)-15 and (±)-5-epi-15, which waschromatographically separated. Intermediate (±)-15 was then transformedinto (±)-V5 through hydrogenolysis in the presence of palladium black(scheme 3). (±)-V6 was obtained from (±)-5-epi-15 accordingly (scheme3).

Example 2—General Methods and Materials

All reactions were carried out under an argon atmosphere unlessotherwise noted. Methylene chloride, tetrahydrofuran, toluene, methanol,dimethylformamide, acetonitrile, diisopropylamine, and triethylaminewere dried prior to use by passage through an activated alumina columnunless otherwise noted (Pangborn, et al., 1996). Anhydrous acetone,ethyl acetate, and 1,2-dichloro-ethane were purchased from commercialsuppliers and stored under argon. Yields refer to chromatographicallyand spectroscopically (¹H NMR) homogenous material, unless otherwisestated.

Reactions were monitored by thin-layer chromatography (TLC) carried outon 0.25 mm E. Merck silica gel plates (60F-254) and were visualizedusing UV light and an ethanolic solution of phosphomolybdic acid andcerium sulfate or an aqueous solution of potassium permanganate. Flashcolumn chromatography using E. Merck silica gel (60, particle size0.040-0.063 mm) was performed as described by Still, et al. (1978). NMRspectra were recorded on a Bruker DRX-600 equipped with a 5 mm DCHcryoprobe, Bruker DRX-500, Bruker AV-400, or Varian NOVA-400 instrumentand calibrated using residual undeuterated solvent for ¹H NMR[δ_(H)=7.26 (CHCl₃), 7.16 (C₆D₅H), 2.05 (D₅H-acetone), 2.50 (D₅H-DMSO),and 5.32 (CDHCl₂) ppm] and ¹³C deuterated solvent for ¹³C NMR[δ_(C)=77.16 (CDCl₃), 128.06 (C₆D₆), 206.68 (d₆-acetone), and 53.84(CD₂Cl₂) ppm] as an internal reference at 298 K (Fulmer, et al., 2010).The following abbreviations were used to designate the multiplicities:s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, b=broad,ap=apparent.

ATR-Infrared (IR) spectra were recorded on a Perkin-Elmer 100 seriesFT-IR spectrometer. High-resolution mass spectra (HRMS) were recorded onan Agilent LC/MSD/TOF mass spectrometer using ESI (electrosprayionization) or a Shimadzu Ion Trap-TOF using ESI. Melting points wererecorded on a Fisher-Johns 12-144 melting point apparatus or a ThomsonHoover uni-melt capillary melting point apparatus. X-Raycrystallographic structures were collected using a Bruker Smart-APEXinstrument (CCD detector) or a Bruker Kappa APEX-II instrument (CCDdetector).

Preparative HPLC separations were performed using a Waters 2767 prep LCsystem equipped with a Waters Atlantis prep T3 OBD column (16×150 mm, 5μm particle size) and monitored using a Waters 2996 photodiode arraydetector.

Example 3—Compound Characterization

Viridicatumtoxin Analog V2:

To a stirred solution of (±)-11 (30 mg, 0.048 mmol, 1.0 equiv) in1,4-dioxane:MeOH (4 mL, 1:1) was added Pd black (28 mg, 0.27 mmol, 5.6equiv) under argon. The flask headspace was exchanged for H₂, and thereaction mixture was stirred for 10 minutes at room temperature. Theflask headspace was re-exchanged for argon, and the mixture was filteredthrough Celite® and concentrated (crude product: 30 mg). A portion ofthis material was purified by reverse-phase prep-HPLC [Waters Atlantisprep T3 OBD, 16×150 mm, 5 μm particle size, 20 mL/min, 50% MeCN in H₂O(0→15 min), then ramp to 70% MeCN (15→25 min), 0.07% TFA buffer, λ=288nm]: t_(R)=17.59 min, to provide pure (±)-V2 (9.5 mg, 17 μmol). (±)-V2:R_(f)=0.1 (silica gel, MeOH:CH₂Cl₂ 1:19); FT-IR (neat) ν_(max)=3435,2921, 1685, 1623, 1585, 1492, 1448, 1399, 1281, 1259, 1198, 1162, 1133,1069, 904, 733 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=18.21 (s, 1H), 14.49 (s,1H), 9.02 (s, 1H), 8.71 (s, 1H), 6.79 (s, 1H), 5.73 (s, 1H), 5.50 (s,1H), 4.82 (s, 1H), 3.88 (s, 3H), 3.71 (d, J=20.1 Hz, 1H), 3.56 (t, J=4.4Hz, 1H), 3.35 (dd, J=18.0, 3.9 Hz, 1H), 3.20 (d, J=20.1 Hz, 1H), 3.15(dd, J=18.0, 5.0 Hz, 1H), 2.20 (m, 1H), 2.04 (m, 1H), 1.81 (td, J=12.6,6.1 Hz, 1H), 1.45 (s, 3H), 1.32 (dd, J=12.6, 6.1 Hz, 1H), 0.90 (s, 3H),0.34 (s, 3H) ppm; ¹³C NMR (CDCl₃, 151 MHz) δ=196.6, 196.5, 192.6, 189.8,173.7, 163.5, 160.9, 158.1, 146.7, 143.6, 135.9, 127.0, 121.7, 120.7,106.9, 106.9, 102.4, 99.8, 77.4, 60.4, 55.8, 50.4, 45.6, 38.5, 34.1,29.3, 25.5, 24.3, 22.9, 21.0 ppm; HRMS (ESI) calcd for C₃₀H₂₉NO₉H⁺[M+H⁺]548.1915. found 548.1902.

Viridicatumtoxin Analog V3:

To a stirred solution of (±)-15-epi-11 (44 mg, 0.057 mmol, 1.0 equiv) in1,4-dioxane:MeOH (5 mL, 1:1) was added Pd black (42 mg, 0.40 mmol, 7equiv) under argon. The flask headspace was exchanged for H₂, and thereaction mixture was stirred for 10 minutes at room temperature. Theflask headspace was re-exchanged for argon, and the mixture was filteredthrough Celite® and concentrated (crude product: 40 mg). A portion ofthis material was purified by reverse-phase prep-HPLC [Waters Atlantisprep T3 OBD, 16×150 mm, 5 μm particle size, 20 mL/min, 50% MeCN in H₂O(0→15 min), then ramp to 70% MeCN (15→25 min), 0.07% TFA buffer, λ=288nm]: t_(R)=17.51 min, to provide pure (±)-V3 (11 mg, 20 μmol). (±)-V3:R_(f)=0.1 (silica gel, MeOH:CH₂Cl₂ 1:19); FT-IR (neat) ν_(max)=3414,2923, 1681, 1622, 1585, 1452, 1401, 1283, 1261, 1203, 1134, 1070, 909,730 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=18.22 (s, 1H), 14.60 (s, 1H), 9.05(s, 1H), 8.74 (s, 1H), 6.79 (s, 1H), 5.73 (s, 1H), 5.49 (s, 1H), 5.47(bs, 1H), 3.88 (s, 3H), 3.85 (m, 1H), 3.44 (t, J=4.6 Hz, 1H), 3.36 (dd,J=18.1, 2.3 Hz, 1H), 3.14 (dd, J=18.1, 5.0 Hz, 1H), 3.08 (d, J=20.0 Hz,1H), 2.19 (m, 1H), 2.04 (m, 1H), 1.83 (td, J=12.6, 6.2 Hz, 1H), 1.41 (s,3H), 1.36 (dd, J=12.6, 5.9 Hz, 1H), 0.91 (s, 3H), 0.39 (s, 3H) ppm; ¹³CNMR (CDCl₃, 151 MHz) δ=196.5, 196.3, 192.6, 189.8, 173.6, 164.0, 161.0,158.1, 146.9, 143.6, 135.9, 127.1, 121.8, 120.6, 107.0, 106.9, 102.4,99.9, 77.3, 60.4, 55.8, 50.5, 45.4, 38.5, 34.1, 29.5, 25.5, 24.6, 23.0,20.8 ppm; HRMS (ESI) calcd for C₃₀H₂₉NO₉H⁺[M+H⁺] 548.1915. found548.1894.

Viridicatumtoxin Analog V4:

To a stirred solution of (±)-15-epi-14 (40 mg, 0.058 mmol, 1.0 equiv) in1,4-dioxane:MeOH (5 mL, 1:1) was added Pd black (43 mg, 0.40 mmol, 7equiv) under argon. The flask headspace was exchanged for H₂, and thereaction mixture was stirred for 10 minutes at room temperature. Theflask headspace was re-exchanged for argon, and the mixture was filteredthrough Celite® and concentrated (crude product: 44 mg). A portion ofthis material was purified by reverse-phase prep-HPLC [Waters Atlantisprep T3 OBD, 16×150 mm, 5 μm particle size, 20 mL/min, 50% MeCN in H₂O(0→15 min), then ramp to 70% MeCN (15→25 min), 0.07% TFA buffer, λ=288nm]: t_(R)=14.25 min, to provide pure (±)-V4 (8.8 mg, 16 μmol). (±V4:R_(f)=0.1 (silica gel, MeOH:CH₂Cl₂ 1:19); FT-IR (neat) ν_(max)=3400,2939, 1683, 1585, 1466, 1400, 1342, 1285, 1216, 1187, 1163, 1132, 1068,914, 824, 731 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=18.19 (s, 1H), 14.20 (s,1H), 9.06 (s, 1H), 6.72 (s, 1H), 5.72 (s, 1H), 5.49 (s, 1H), 4.93 (s,1H), 4.09 (s, 3H), 3.94 (s, 3H), 3.86 (d, J=19.9 Hz, 1H), 3.44 (t, J=4.5Hz, 1H), 3.36 (dd, J=18.1, 4.0 Hz, 1H), 3.13 (dd, J=18.1, 5.1 Hz, 1H),3.08 (d, J=19.9 Hz, 1H), 2.20 (m, 1H), 2.05 (m, 1H), 1.85 (td, J=12.5,6.1 Hz, 1H), 1.42 (s, 3H), 1.34 (dd, J=12.5, 6.2 Hz, 1H), 0.91 (s, 3H),0.37 (s, 3H) ppm; ¹³C NMR (CDCl₃, 151 MHz) δ=196.2, 196.1, 193.1, 190.0,173.6, 165.0, 160.4, 159.6, 148.1, 142.1, 136.2, 126.9, 121.6, 121.5,110.0, 107.9, 99.9, 98.1, 77.2, 59.7, 56.8, 55.6, 50.5, 45.2, 38.5,34.0, 29.4, 25.5, 24.5, 23.0, 20.8 ppm; HRMS (ESI) calcd forC₃₁H₃₁NO₉H⁺[M+H⁺] 562.2072. found 562.2082.

(±)-15 and (±)-5-epi-15:

This reaction was run twice in parallel. To two separate batches ofsubstrate (±)-11 (25 mg each, 0.032 mmol each, 1.0 equiv each) was addedsolid NaCNBH₃ (8 mg each, 0.13 mmol each, 4.0 equiv each). Then, AcOH(2.5 mL each) was rapidly injected into the reaction vessel, and themixture was vigorously stirred at room temperature for 25 minutes. Thetwo reaction mixtures were combined for work up by pouring them intowater (25 mL). Brine (5 mL) was added, and the mixture was extractedwith EtOAc (35 mL). The organic phase was washed with additional brine(2×25 mL), and the organic layer was dried over Na₂SO₄, filtered andconcentrated. The crude residue was purified by preparative TLC (silicagel, 10% acetone:toluene) to provide (±)-15 (26 mg, not pure) and isomer(±)-5-epi-15 (13 mg, 0.017 mmol, 27%, yellow foam). Product (±)-15 wasfurther purified by preparative TLC (silica gel, 5% EtOAc:CH₂Cl₂) togive pure compound (±)-15 (15 mg, 0.020 mmol, 31%) as a yellow foam.

(±)-15:

R_(f)=0.6 (silica gel, acetone:toluene 3:17); FT-IR (neat) ν_(max)=3397,2916, 1702, 1591, 1512, 1481, 1448, 1405, 1371, 1339, 1319, 1254, 1195,1143, 1109, 1081, 1032, 986, 914, 812, 734, 695 cm⁻¹; ¹H NMR (C₆D₆, 500MHz) δ=15.00 (s, 1H), 7.58-7.55 (m, 2H), 7.28-7.25 (m, 2H), 7.24-7.20(m, 2H), 7.12 (m, 1H), 7.05-6.98 (m, 3H), 6.26 (s, 1H), 5.56 (s, 1H),5.14 (d, J=12.2 Hz, 1H, AB system), 5.10 (d, J=12.2 Hz, 1H, AB system),4.89 (ap s, 3H), 4.34 (d, J=8.6 Hz, 1H), 3.56 (d, J=17.6 Hz, 1H), 3.26(s, 3H), 2.85 (s, 3H), 2.83 (m, 1H), 2.77-2.64 (m, 3H), 2.31 (m, 1H),2.04 (m, 1H), 1.91 (ddd, J=12.3, 12.3, 6.1 Hz, 1H), 1.65 (s, 3H), 1.37(dd, J=13.2, 6.0 Hz, 1H), 1.15 (s, 3H), 0.69 (s, 3H) ppm; ¹³C NMR (C₆D₆,126 MHz) δ=197.0, 185.4, 180.3, 168.6, 166.3, 159.7, 158.8, 149.2,137.9, 137.2, 135.7, 132.5, 128.9, 128.7, 128.6, 128.51, 128.48, 127.2,127.0, 122.5, 121.1, 109.2, 109.0, 106.2, 97.5, 78.6, 75.3, 72.3, 71.2,59.3, 54.9, 54.7, 42.8, 42.6, 38.7, 34.4, 26.1, 24.6, 23.4, 22.7, 21.5ppm; HRMS (ESI) calcd for C₄₅H₄₃NO₉Na⁺[M+Na⁺] 764.2830. found 764.2853.

(±)-5-epi-15:

R_(f)=0.7 (silica gel, acetone:toluene 3:17); FT-IR (neat) ν_(max)=3447,2917, 1708, 1593, 1514, 1485, 1449, 1407, 1373, 1345, 1317, 1259, 1192,1138, 1115, 1086, 994, 913, 813, 735, 696 cm⁻¹; ¹H NMR (C₆D₆, 500 MHz)*δ=15.33 (s, 1H), 7.57-7.54 (m, 2H), 7.35-7.32 (m, 2H), 7.22-7.18 (m,2H), 7.12-7.06 (m, 3H), 7.04 (m, 1H), 6.25 (s, 1H), 5.59 (s, 1H), 5.25(d, J=12.1 Hz, 1H, AB system), 5.13 (d, J=12.1 Hz, 1H, AB system), 4.91(s, 2H), 4.75 (bs, 1H), 3.44 (bs, 1H), 3.36 (m, 1H), 3.24 (s, 3H), 2.91(bs, 3H), 2.64-2.56 (m, 2H), 2.38-2.25 (m, 2H), 2.10-1.97 (m, 2H), 1.78(s, 3H), 1.38 (dd, J=12.5, 6.0 Hz, 1H), 1.12 (s, 3H), 0.59 (s, 3H) ppm;¹³C NMR (C₆D₆, 126 MHz)* δ=167.5, 159.7, 158.9, 138.0, 137.3, 135.8,132.5, 128.9, 128.7, 128.63, 128.56, 127.1, 126.4, 122.5, 121.2, 108.6,106.7, 97.5, 72.4, 71.2, 59.1, 56.3, 54.8, 38.5, 34.4, 26.0, 24.3, 23.5,21.5 ppm; HRMS (ESI) calcd for C₄₅H₄₃NO₉Na⁺[M+Na⁺] 764.2830. found764.2832.

*Due to signal broadening, 1 proton signal and 12 carbon signals couldnot be identified.

Viridicatumtoxin Analog V5:

Following conditions similar to those of Stork, et al. (1996), Pd black(12 mg, 0.11 mmol, 4.1 equiv) was added to a stirred solution of (±)-15(20 mg, 0.027 mmol, 1.0 equiv) in THF:MeOH (1.2 mL, 1:1) under argon.The flask headspace was exchanged for H₂, and the reaction mixture wasstirred for 10 minutes at room temperature. The flask headspace wasre-exchanged for argon, and the mixture was filtered through cotton andconcentrated to give (±)-V5 (15 mg, 0.026 mmol, 96%) as a yellow powder.(±)-V5: R_(f) ⁼0.1 (silica gel, MeOH:CH₂Cl₂ 1:19); FT-IR (neat)ν_(max)=3399, 2917, 1625, 1595, 1490, 1471, 1451, 1401, 1309, 1273,1200, 1084, 909, 732 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=18.00 (s, 1H),15.12 (s, 1H), 9.25 (bs, 1H), 8.80 (s, 1H), 6.63 (s, 1H), 5.91 (bs, 1H),5.49 (bs, 1 H), 5.16 (bs, 1H), 4.39 (d, J=4.5 Hz, 1H), 3.85 (s, 3H),3.57 (s, 3H), 3.36 (d, J=17.5 Hz, 1H), 3.13 (m, 1H), 2.88 (d, J=17.5 Hz,1H), 2.85 (dd, J=18.9, 5.4 Hz, 1H), 2.58 (dd, J=18.9, 10.0 Hz, 1H), 2.22(m, 1H), 2.04 (m, 1H), 1.77 (ddd, J=12.7, 12.7, 6.1 Hz, 1H), 1.48 (s,3H), 1.38 (dd, J=12.7, 6.2 Hz, 1H), 0.93 (s, 3H), 0.50 (s, 3H) ppm; ¹³CNMR (CDCl₃, 151 MHz) δ=195.0, 194.0, 191.5, 173.2, 166.1, 160.5, 158.2,147.6, 137.1, 135.7, 123.5, 122.5, 121.2, 106.4, 105.7, 100.6, 99.7,76.7, 76.5, 60.0, 57.1, 55.6, 41.9, 38.4, 38.0, 34.2, 32.8, 25.8, 24.3,23.0, 21.1 ppm; HRMS (ESI) calcd for C₃₁H₃₃NO₉Na⁺[M+Na⁺] 586.2048. found586.2045.

Viridicatumtoxin Analog V6:

Following conditions similar to those of Stork, et al., (1996), Pd black(8.6 mg, 0.081 mmol, 4.5 equiv) was added to a stirred solution of(±)-5-epi-15 (13 mg, 0.018 mmol, 1.0 equiv) in THF:MeOH (1.0 mL, 1:1)under argon. The flask headspace was exchanged for H_(z), and thereaction mixture was stirred for 10 minutes at room temperature. Theflask headspace was re-exchanged for argon, and the mixture was filteredthrough cotton and concentrated to give (±)-V6 (10 mg, 0.018 mmol,quant.) as a yellow powder. (±)-V6: R_(f)=0.1 (silica gel, MeOH:CH₂Cl₂1:19); FT-IR (neat) ν_(max)=3419, 2921, 1626, 1594, 1490, 1471, 1450,1404, 1301, 1201, 1139, 1088, 907, 733 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz)δ=17.94 (s, 1H), 15.34 (s, 1H), 9.11 (s, 1H), 8.84 (s, 1H), 6.60 (s,1H), 5.85 (s, 1H), 5.49 (s, 1H), 5.00 (s, 1H), 4.86 (s, 1H), 3.85 (s,3H), 3.52 (bs, 3H), 3.22 (d, J=17.9 Hz, 1H), 3.13 (d, J=17.9 Hz, 1H),3.10 (m, 1H), 2.92 (m, 1H), 2.60 (dd, J=19.6, 10.9 Hz, 1H), 2.22 (m,1H), 2.03 (m, 1H), 1.82 (td, J=12.6, 6.1 Hz, 1H), 1.54 (s, 3H), 1.32(dd, J=12.6, 6.1 Hz, 1H), 0.89 (s, 3H), 0.42 (s, 3H) ppm; ¹³C NMR(CDCl₃, 151 MHz) δ=195.2, 194.8 (b), 192.1, 173.0, 166.6, 160.6, 158.1,148.2, 137.6, 134.2, 124.2, 122.1, 120.8, 106.9, 105.3, 100.7, 99.2,76.6, 75.6, 59.4, 57.0, 55.6, 43.9, 39.2 (b), 38.3, 34.1, 30.8 (b),25.7, 24.1, 23.0, 21.2 ppm; HRMS (ESI) calcd for C₃₁H₃₃NO₉Na⁺[M+Na⁺]586.2048. found 586.2036.

Example 4: Biological Activity A. Bacterial Strains and Growth Media:

Four clinical strains were used for these studies, Enterococcus faecalisS613, Enterococcus faecium isolate 105, Methicillin-ResistantStaphylococcus aureus 371 (MRSA 371) and Acinetobacter baumannii AB210.Enterococcus strains were cultured in 80% Lysogeny Broth (LB) & 20%Brain Heart Infusion (BHI). MRSA 371 and AB210 were cultured in 100% LB.

B. Minimal Inhibitory Concentration (MIC) Assays:

Micro-broth MIC assays were performed in triplicate using 96-wellplates. Wells were filled with 150 μL of broth media and inoculated with2 μL of stationary phase culture. The concentration of the testantibiotics increased in 2-fold increments and ranged from 0.25-128μg/mL. Plates were incubated overnight at 37° C. and the MICs weredefined as the lowest drug concentration that had no growth after 16-24hours.

C. Biological Evaluation of Analogs

The minimum inhibitory concentrations against both selectedGram-positive and Gram-negative strains were determined for analogsV2-V6 and the results are shown in table 1.

TABLE 1 Minimum Inhibitory Concentration (MIC) data of compounds againstGram-positive and Gram-negative bacteria and comparison with selectedliterature data^(a) Gram-(+) Gram-(−) E. faecalis E. faecium MRSA A.baumannii S613 501 371 AB210 (±)-1 1 0.5 4 64 (+)-2 1 1 4 64 (±)-V2 0.50.5 2 64 (±)-V3 4 2 8 64 (±)-V4 4 4 4 64 (±)-V5 1 1 8 64 (±)-V6 0.5 0.52 64 ^(a)MIC assays were run in triplicate; data are given in units ofμg/mL.

All of the compositions and/or methods disclosed and claimed herein canbe made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions and/or methods and in the steps or in the sequence ofsteps of the method described herein without departing from the concept,spirit and scope of the invention. More specifically, it will beapparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the spirit, scope and concept of theinvention as defined by the appended claims.

V. REFERENCES

The following references, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference:

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What is claimed is:
 1. A compound of the formula:

wherein: X₁ is absent such that atoms 16 and 17 are only connected bythe shown single bond, a covalent bond such that a double bond is formedbetween atoms 16 and 17, —O—, alkanediyl_((C≦8)), or substitutedalkanediyl_((C≦8)); Y₁, Y₂, and Y₃ are each independently alkyl_((C≦12))or substituted alkyl_((C≦12)); R₁ is hydrogen, hydroxy, alkoxy_((C≦8)),substituted alkoxy_((C≦8)), or oxo, provided that when R₁ is oxo, thebond between R₁ and atom number 5 is a double bond and when the bondbetween R₁ and atom number 5 is a double bond then R₁ is oxo; R₂ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₃, R₆, R₂, and R₁₀ are each independently selected from: hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; R₄ and R₅are each independently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 2. The compound of claim 1 furtherdefined as:

wherein: X₁ is absent such that atoms 16 and 17 are only connected bythe shown single bond, a covalent bond such that a double bond is formedbetween atoms 16 and 17, —O—, alkanediyl_((C≦8)), or substitutedalkanediyl_((C≦8)); Y₁, Y₂, and Y₃ are each independently alkyl_((C≦12))or substituted alkyl_((C≦12)); R₁ is hydrogen, hydroxy, alkoxy_((C≦8)),substituted alkoxy_((C≦8)), or oxo, provided that when R₁ is oxo, thebond between R₁ and atom number 5 is a double bond and when the bondbetween R₁ and atom number 5 is a double bond then R₁ is oxo; R₂ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ and R₅ are each independently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 3. The compound of either claim 1or claim 2 further defined as:

wherein: X₁ is a covalent bond such that a double bond is formed betweenatoms 16 and 17, —O—, alkanediyl_((C≦8)), or substitutedalkanediyl_((C≦8)); R₁ is hydrogen, hydroxy, alkoxy_((C≦8)), substitutedalkoxy_((C≦8)), or oxo, provided that when R₁ is oxo, the bond betweenR₁ and atom number 5 is a double bond and when the bond between R₁ andatom number 5 is a double bond then R₁ is oxo; R₂ is hydrogen, amino,carboxy, cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)),alkoxy_((C≦12)), alkylamino_((C≦12)), dialkylamino_((C≦18)), or asubstituted version of any of these groups; R₄ and R₅ are eachindependently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 4. The compound according to anyone of claims 1-3 further defined as:

wherein: R₁ is hydrogen, hydroxy, alkoxy_((C≦8)), substitutedalkoxy_((C≦8)), or oxo, provided that when R₁ is oxo, the bond betweenR₁ and atom number 5 is a double bond and when the bond between R₁ andatom number 5 is a double bond then R₁ is oxo; R₂ is hydrogen, amino,carboxy, cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)),alkoxy_((C≦12)), alkylamino_((C≦12)), dialkylamino_((C≦18)), or asubstituted version of any of these groups; R₄ and R₅ are eachindependently selected from: hydrogen, alkyl_((C≦8)),alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 5. The compound according to anyone of claims 1-4 further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein the biomolecule is a protein, a polypeptide,an amino acid, a cofactor, an imaging agent, an antibody, a fatty acid,a nucleic acid, or a small molecule therapeutic agent; and R₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 6. The compound according to anyone of claims 1-5 further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein: the linker is alkanediyl_((C≦12)),alkenediyl_((C≦12)), arenediyl_((C≦12)), heteroarenediyl_((C≦12)),heterocycloalkanediyl_((C≦12)) or a substituted version of any of thesegroups; and the biomolecule is a protein, a polypeptide, an antibody, animaging agent, or a small molecule therapeutic agent; R₉ is hydrogen,amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)),alkoxy_((C≦12)), alkylamino_((C≦12)), dialkylamino_((C≦18)),amido_((C≦12)), or a substituted version of any of these groups; or—NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen, alkyl_((C≦8)), orsubstituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8)) or substitutedalkanediyl_((C≦8)); and R₁₄ and R₁₅ are each independently selectedfrom: hydrogen, alkyl_((C≦12)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦12)), or a substituted version of any of these groups; or R₁₄and R₁₅ are taken together and are alkanediyl_((C≦8)),alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or a substituted version ofany of these groups; or a pharmaceutically acceptable salt or tautomerthereof.
 7. The compound according to any one of claims 1-6 furtherdefined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), substitutedalkyl_((C≦8)), or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12)) orsubstituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule wherein: the linker is alkanediyl_((C≦12)),alkenediyl_((C≦12)), arenediyl_((C≦12)), heteroarenediyl_((C≦12)),heterocycloalkanediyl_((C≦12)) or a substituted version of any of thesegroups; and the biomolecule is an antibody, an imaging agent, a protein,or a small molecule therapeutic agent; R₉ is hydrogen, amino, carboxy,cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), amido_((C≦12)), or asubstituted version of any of these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅;wherein: R₁₂ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8));R₁₃ is alkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ andR₁₅ are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 8. The compound according to anyone of claims 1-7 further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen or -linker-biomolecule; wherein: thelinker is alkanediyl_((C≦12)), alkenediyl_((C≦12)), arenediyl_((C≦12)),heteroarenediyl_((C≦12)), or heterocycloalkanediyl_((C≦12)); and thebiomolecule is an antibody; and R₉ is hydrogen, amino, carboxy, cyano,halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), amido_((C≦12)), or asubstituted version of any of these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅;wherein: R₁₂ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8));R₁₃ is alkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ andR₁₅ are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 9. The compound according to anyone of claims 1-8 further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, halo, or hydroxy; alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule; and R₉ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), amido_((C≦12)), or asubstituted version of any of these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅;wherein: R₁₂ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8));R₁₃ is alkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ andR₁₅ are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 10. The compound according to anyone of claims 1-5 further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, halo, or hydroxy; alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule; and R₉ is hydrogen, amino, cyano, halo, hydroxy;alkoxy_((C≦12)), amido_((C≦12)), or a substituted version of either ofthese groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; or a pharmaceuticallyacceptable salt or tautomer thereof.
 11. The compound according to anyone of claims 1-5 and 10 further defined as:

wherein: R₁ is hydroxy, alkoxy_((C≦8)), or substituted alkoxy_((C≦8));R₂ is hydrogen, amino, halo, or hydroxy; alkylamino_((C≦12)),dialkylamino_((C≦18)), or a substituted version of any of these groups;R₄ is hydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)), R₅ ishydrogen, alkyl_((C≦8)), alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)),alkanediyl_((C≦8))-heteroaryl_((C≦8)),alkanediyl_((C≦8))-alkylamino_((C≦8)),alkanediyl_((C≦8))-dialkylamino_((C≦8)), or a substituted version of anyof these groups; R₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; or a-linker-biomolecule; and R₉ is —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ ishydrogen, alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ isalkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅are each independently selected from: hydrogen, alkyl_((C≦12)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦12)), or a substituted versionof any of these groups; or a pharmaceutically acceptable salt ortautomer thereof.
 12. The compound according to any one of claims 1-11,wherein R₁ is alkoxy_((C≦8)) or substituted alkoxy_((C≦8)).
 13. Thecompound of claim 12, wherein R₁ is alkoxy_((C≦6)).
 14. The compound ofeither claim 12 or claim 13, wherein R₁ is methoxy.
 15. The compoundaccording to any one of claims 1-4, wherein R₁ is oxo.
 16. The compoundaccording to any one of claims 1-15, wherein R₂ is hydrogen.
 17. Thecompound according to any one of claims 1-15, wherein R₂ is halo. 18.The compound of claim 17, wherein R₂ is fluoro, chloro, bromo, or iodo.19. The compound according to any one of claims 1-15, wherein R₂ ishydroxy.
 20. The compound according to any one of claims 1-15, whereinR₂ is amino.
 21. The compound according to any one of claims 1-15,wherein R₂ is dialkylamino_((C≦12)) or substituteddialkylamino_((C≦12)).
 22. The compound of claim 21, wherein R₂ isdialkylamino_((C≦12)).
 23. The compound of either claim 21 or claim 22,wherein R₂ is dimethylamino or (2-aminoethyl)methylamino.
 24. Thecompound according to any one of claims 1-23, wherein R₄ is hydrogen.25. The compound according to any one of claims 1-24, wherein R₅ ishydrogen.
 26. The compound according to any one of claims 1-24, whereinR₅ is alkanediyl_((C≦8))-heterocycloalkyl_((C≦8)).
 27. The compound ofclaim 26, wherein R₅ is CH₂CH₂N(CH₂)₄.
 28. The compound according to anyone of claims 1-24, wherein R₅ is alkanediyl_((C≦8))-alkylamino_((C≦8)).29. The compound of claim 28, wherein R₅ is (CH₂)₂NH(CH₂)₄CH(NH₂)CO₂H.30. The compound according to any one of claims 1-29, wherein R₈ ishydrogen.
 31. The compound according to any one of claims 1-29, whereinR₈ is —X₂—R₁₁.
 32. The compound of claim 31, wherein X₂ isalkanediyl_((C≦6)) or substituted alkanediyl_((C≦6)).
 33. The compoundof either claim 31 or claim 32, wherein X₂ is alkanediyl_((C≦6)). 34.The compound according to any one of claims 31-33, wherein X₂ is—CH₂CH₂CH₂— or —CH₂CH₂—.
 35. The compound according to any one of claims31-34, wherein R₁₁ is amino.
 36. The compound according to any one ofclaims 31-34, wherein R₁₁ is heterocycloalkyl_((C≦12)), or substitutedheterocycloalkyl_((C≦12)).
 37. The compound of claim 36, wherein R₁₁ is4-N-methyl-piperazinyl.
 38. The compound of according to any one ofclaims 1-29, wherein R₈ is a -linker-biomolecule.
 39. The compound ofclaim 38, wherein the linker is alkanediyl_((C≦12)),alkenediyl_((C≦12)), arenediyl_((C≦12)), heteroarenediyl_((C≦12)),heterocycloalkanediyl_((C≦12)) or a substituted version of any of thesegroups.
 40. The compound of either claim 38 or claim 39, wherein thebiomolecule is an antibody, a protein, or a small molecule therapeuticagent.
 41. The compound of claim 40, wherein the biomolecule is anantibody.
 42. The compound according to any one of claims 1-10 and12-41, wherein R₉ is hydrogen.
 43. The compound according to any one ofclaims 1-10 and 12-41, wherein R₉ is halo.
 44. The compound of claim 43,wherein R₉ is fluoro or iodo.
 45. The compound according to any one ofclaims 1-10 and 12-41, wherein R₉ is amido_((C≦12)) or substitutedamido_((C≦12)).
 46. The compound of claim 45, wherein R₉ is—NHC(O)CH₂NH₂.
 47. The compound according to any one of claims 1-41,wherein R₉ is —NR₁₂C(O)R₁₃—NR₁₄R₁₅.
 48. The compound of claim 47,wherein R₁₂ is hydrogen.
 49. The compound of claim 47, wherein R₁₂ isalkyl_((C≦6)) or substituted alkyl_((C≦6)).
 50. The compound accordingto any one of claims 47-49, wherein R₁₃ is alkanediyl_((C≦8)).
 51. Thecompound of claim 50, wherein R₁₃ is —CH₂—.
 52. The compound accordingto any one of claims 47-51, wherein R₁₄ is alkyl_((C≦12)) or substitutedalkyl_((C≦12)).
 53. The compound of claim 52, wherein R₁₄ is t-butyl.54. The compound according to any one of claims 47-53, wherein R₁₅ ishydrogen.
 55. The compound according to any one of claims 1-54, whereinthe compound is further defined as:

or a pharmaceutically acceptable salt, tautomer, or optical isomerthereof.
 56. A pharmaceutical composition comprising a compoundaccording to any one of claims 1-55 and an excipient.
 57. Thecomposition of claim 56, wherein the composition is formulated foradministration: orally, intraadiposally, intraarterially,intraarticularly, intracranially, intradermally, intralesionally,intramuscularly, intranasally, intraocularly, intrapericardially,intraperitoneally, intrapleurally, intraprostatically, intrarectally,intrathecally, intratracheally, intratumorally, intraumbilically,intravaginally, intravenously, intravesicularlly, intravitreally,liposomally, locally, mucosally, parenterally, rectally,subconjunctival, subcutaneously, sublingually, topically, transbuccally,transdermally, vaginally, in crmes, in lipid compositions, via acatheter, via a lavage, via continuous infusion, via infusion, viainhalation, via injection, via local delivery, or via localizedperfusion.
 58. The composition of either claim 56 or claim 57, whereinthe composition is formulated for administration: orally, intravenously,or topically.
 59. A method of treating a disease or disorder comprisingadministering a pharmaceutically effective amount of a compound orcomposition according to any one of claims 1-58.
 60. The method of claim59, wherein the disease or disorder is a microbial infection.
 61. Themethod of either claim 59 or claim 60, wherein the microbial infectionis a bacterial infection.
 62. The method according to any one of claims59-61, wherein the infection is by a gram positive or gram negativebacteria.
 63. The method according to any one of claims 59-62, whereinthe disease is a bacteria infection by Enterococcus faecalis,Enterococcus faecium, Staphylococcus aureus, Acinetobacter baumannii,Escherichia coli, Acinetobacter calcoaceticus, Staphycococcusepidermidis, Pseudomonas aeruginosa, Klebsiella aerogenes, Candidaalbicans, Salmonella typhinurium, Streptococcus pneumoniae, Micrococcusluteus, Bacillus cerues, or Bacillus subtilis.
 64. The method of claim63, wherein the disease is a bacteria infection by Staphylococcus aureus503, Staphylococcus aureus 209, Staphylococcus aureus RN420,Methicillin-resistant Staphylococcus aureus CCARM 3167,Methicillin-resistant Staphylococcus aureus 371, Methicillin-resistantStaphylococcus aureus CCARM 3506, quinolone-resistant Staphylococcusaureus CCARM 3505, quinolone-resistant Staphylococcus aureus CCARM 3519,Bacillus subtilis KCTC 1021, Bacillus cerues KCTC 1661, Micrococcusluteus KCTC 1056, Streptococcus pneumoniae KCTC 3932, Streptococcuspneumoniae KCTC 5412, Enterococcus faecium 501, Enterococcus faeciumKCTC 3122, Enterococcus faecalis 5613, Enterococcus faecalis KCTC 5191,Enterococcus faecalis KCTC 3511, Staphycococcus epidermidis KCTC 3958,Salmonella typhinurium KCTC 1926, Acinetobacter calcoaceticus KCTC 2357,Escherichia coli CCARM 1358, Escherichia coli KCTC 1682, Pseudomonasaeruginosa KCTC 2004, Pseudomonas aeruginosa KCTC 2742, Klebsiellaaerogenes KCTC 2619, Acinetobacter baumannii AB210, or Candida albicansKCTC
 7535. 65. The method according to any one of claim 61-64, whereinthe bacteria is a drug-resistant bacteria.
 66. The method according toany one of claims 59-65 further comprising administering a secondtherapeutic agent.
 67. The method of claim 66, wherein the secondtherapeutic agent is an antibiotic.
 68. The method of either claim 66 orclaim 67, wherein the second therapeutic agent is a tetracyclineantibiotic.
 69. The method according to any one of claims 66-68, whereinthe second therapeutic agent is viridicatumtoxin A, viridicatumtoxin B,vancomycin, tetracycline, spirohexaline, minocycline, tigecycline,doxycycline, a β-lactam antibiotic, an aminoglycoside antibiotic, asulfonamide antibiotic, a macrolide antibiotic, a glycopeptideantibioitic, an ansamycin antibiotic, an oxazolidinone antibiotic, aquinolone antibiotic, a streptogramin antibiotic, or a lipopeptideantibiotic.
 70. The method of either claim 59 or claim 60, wherein themicrobial infection is a viral infection.
 71. The method according toany one of claims 59, 60, and 70, wherein the virus is a poxvirus. 72.The method of claim 71, wherein the poxvirus is variola virus, vacciniavirus, or molluscum contagiosum.
 73. The method according to any one ofclaims 59, 60, and 70-72 further comprising administering a secondtherapeutic agent.
 74. The method of claim 73, wherein the secondtherapeutic agent is an interferon or antiviral compound.
 75. The methodof claim 59, wherein the disease or disorder is cancer.
 76. The methodof either claim 59 or claim 75, wherein the cancer is a carcinoma,sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma,or seminoma.
 77. The method of either claim 59 or claim 75, wherein thecancer is of the bladder, blood, bone, brain, breast, central nervoussystem, cervix, colon, endometrium, esophagus, gall bladder,gastrointestinal tract, genitalia, genitourinary tract, head, kidney,larynx, liver, lung, muscle tissue, neck, oral or nasal mucosa, ovary,pancreas, prostate, skin, spleen, small intestine, large intestine,stomach, testicle, or thyroid.
 78. The method according to any one ofclaim 59 or 75-77 further comprising administering a second therapeuticagent.
 79. The method of claim 78, wherein the second therapeutic agentis a second chemotherapeutic agent, radiotherapy, immunotherapy, orsurgery.
 80. A method of inhibiting the activity of a bacterial ribosomefor the treatment of a disease or disorder comprising administering acompound or composition according to any one of claims 1-58.
 81. Amethod of inhibiting the activity of a bacterial UPP synthase for thetreatment of a disease or disorder comprising administering a compoundor composition according to any one of claims 1-58.
 82. A method ofpreparing a compound of the formula:

wherein: Y₄, Y₅, and Y₆ are each independently hydrogen, alkyl_((C≦8))or substituted alky_((C≦8)); R₁₆ and R₁₆′ are each independentlyhydrogen, alkoxy_((C≦8)), aralkoxy_((C≦12)), substituted alkoxy_((C≦8)),or substituted aralkoxy_((C≦12)); or R₁₆ and R₁₆′ are taken together andare alkoxydiyl_((C≦12)); R₁₇ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), or a substituted version ofany of these groups; or —C(O)OCH₂CH₂Si(CH₃)₃; X₃ is O, NH, S, or CH₂; X₄is hydroxy, amino, mercapto, O, NH, or S provided that when X₄ is O, NH,or S, the bond to which the atom is attached is a double bond andprovide that when the bond to which the atom is attached is a doublebond, then X₄ is O, NH, or S; X₅ is O, NH, or S; R₁₈ is hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; or—X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12)) or substitutedalkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido, carboxy, orcyano, alkenyl_((C≦6)), alkynyl_((C≦6)), heterocycloalkyl_((C≦12)),alkylamino_((C≦8)), dialkylamino_((C≦8)), alkoxy_((C≦8)), or asubstituted version of any of these groups; R₁₉ is hydrogen, amino,carboxy, cyano, halo, hydroxy, nitro, or sulfo; alkyl_((C≦12)),alkoxy_((C≦12)), alkylamino_((C≦12)), dialkylamino_((C≦18)),amido_((C≦12)), or a substituted version of any of these groups; or—NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen, alkyl_((C≦8)), orsubstituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8)) or substitutedalkanediyl_((C≦8)); and R₁₄ and R₁₅ are each independently selectedfrom: hydrogen, alkyl_((C≦12)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦12)), or a substituted version of any of these groups; or R₁₄and R₁₅ are taken together and are alkanediyl_((C≦8)),alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or a substituted version ofany of these groups; and R₂₀ and R₂₁ are each independently hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; or asalt, tautomer, or optical isomer thereof; comprising reacting acompound of the formula:

wherein: Y₄, Y₅, Y₆, R₁₆, R₁₆′, X₅, R₁₈, R₁₉, and R₂₀ are as definedabove; and X₄ is O, NH, or S; with a compound of the formula:

wherein: X₃ and R₁₇ are as defined above; R₂₁ is hydrogen,alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)),acyl_((C≦8)), or a substituted version of any of these groups; and R₂₂is aryl_((C≦12)), aralkyl_((C≦12)), or a substituted version of eitherof these groups; in the presence of a base.
 83. The method of claim 82,wherein Y₄, Y₅, and Y₆ are alkyl_((C≦6)).
 84. The method of claim 83,wherein Y₄, Y₅, and Y₆ are methyl.
 85. The method according to any oneof claims 82-84, wherein R₁₆ and R₁₆′ are alkoxy_((C≦8)) or substitutedalkoxy_((C≦8)).
 86. The method of claim 85, wherein R₁₆ and R₁₆′ aremethoxy.
 87. The method according to any one of claims 82-86, whereinR₁₇ is hydrogen, amino, halo, hydroxy; alkylamino_((C≦12)), substitutedalkylamino_((C≦12)), dialkylamino_((C≦18)), substituteddialkylamino_((C≦18)), or —C(O)OCH₂CH₂Si(CH₃)₃.
 88. The method of claim87, wherein R₁₇ is amino, hydroxy, or halo.
 89. The method of claim 87,wherein R₁₇ is dimethylamino or 2-aminoethylmethylamino.
 90. The methodaccording to any one of claims 82-89, wherein X₃ and X₅ are O.
 91. Themethod according to any one of claims 82-90, wherein X₄ is hydroxy. 92.The method according to any one of claims 82-90, wherein X₄ is O. 93.The method according to any one of claims 82-92, wherein R₁₈ ishydrogen, alkyl_((C≦8)), substituted alkyl_((C≦8)), or —X₂—R₁₁, wherein:X₂ is alkanediyl_((C≦12)) or substituted alkanediyl_((C≦12)) and R₁₁ ishydroxy, amino, heterocycloalkyl_((C≦12)), substitutedheterocycloalkyl_((C≦12)), alkylamino_((C≦8)), or substitutedalkylamino_((C≦8)),
 94. The method of claim 93, wherein R₁₈ is hydrogen.95. The method according to any one of claims 82-94, wherein R₁₉ ishydrogen, amino, halo, hydroxy, alkylamino_((C≦12)), substitutedalkylamino_((C≦12)), amido_((C≦12)), substituted amido_((C≦12)), or—NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen, R₁₃ isalkanediyl_((C≦8)) or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅are each independently hydrogen, alkyl_((C≦12)), substitutedalkyl_((C≦12)), aryl_((C≦12)), substituted aryl_((C≦12)),aralkyl_((C≦12)), substituted aralkyl_((C≦12)), acyl_((C≦12)), orsubstituted acyl_((C≦12)).
 96. The method of claim 95, wherein R₁₉ ishydrogen, halo, amido_((C≦12)), substituted amido_((C≦12)), or—NHC(O)CH₂—NHR₁₅ wherein R₁₅ is hydrogen, alkyl_((C≦12)), or substitutedalkyl_((C≦12)).
 97. The method according to any one of claims 82-96,wherein R₂₀ is hydrogen, alkyl_((C≦6)), or substituted alkyl_((C≦6)).98. The method of claim 97, wherein R₂₀ is methyl.
 99. The methodaccording to any one of claims 82-98, wherein R₂₁ is aralkyl_((C≦12)) orsubstituted aralkyl_((C≦12)).
 100. The method of claim 99, wherein R₂₁is benzyl.
 101. The method according to any one of claims 82-100,wherein R₂₂ is aryl_((C≦12)) or substituted aryl_((C≦12)).
 102. Themethod of claim 96.3, wherein R₂₂ is phenyl.
 103. The method accordingto any one of claims 82-98, wherein R₁₇ is not hydrogen or—C(O)OCH₂CH₂Si(CH₃)₃, R₁₈ is not benzyl, R₁₉ are not hydrogen, and R₂₀is not methyl.
 104. The method according to any one of claims 82-103,wherein the base is an alkoxide_((C≦18)).
 105. The method of claim 104,wherein the alkoxide_((C≦18)) is t-butoxide.
 106. The method accordingto any one of claims 82-105, wherein the method comprises adding fromabout 1.0 to about 2.0 equivalents of base relative to the compound offormula VII.
 107. The method of claim 106, wherein the method comprisesadding from about 1.0 to about 1.5 equivalents of base.
 108. The methodof either claim 106 or claim 107, wherein the method comprises addingabout 1.2 equivalents of base.
 109. The method according to any one ofclaims 82-108, wherein the method comprises adding from about 0.9 toabout 2.0 equivalents of the compound of formula VIII relative to thecompound of formula VII.
 110. The method of claim 109, wherein themethod comprises adding from about 1.0 to about 1.5 equivalents of thecompound of formula VIII.
 111. The method of either claim 109 or claim110, wherein the method comprises adding about 1.1 equivalents of thecompound of formula VIII.
 112. The method according to any one of claims82-111, wherein the method further comprises a solvent.
 113. The methodof claim 112, wherein the solvent is an organic solvent.
 114. The methodof either claim 112 or claim 113, wherein the solvent is anarene_((C≦12)).
 115. The method of claim 114, wherein the solvent istoluene.
 116. The method according to any one of claims 82-115, whereinthe reaction comprises running the reaction at a temperature from about0° C. to about 50° C.
 117. The method of claim 116, wherein thetemperature is from about 20° C. to about 35° C.
 118. The method ofeither claim 116 or 117, wherein the temperature is about 25° C. 119.The method of claim 116, wherein the temperature is about roomtemperature.
 120. The method according to any one of claims 82-119,wherein the reaction is run for a time period from about 5 minutes toabout 2 hours.
 121. The method of claim 120, wherein the time period isfrom about 10 minutes to about 45 minutes.
 122. The method of eitherclaim 120 or claim 121, wherein the time period is about 15 minutes.123. The method according to any one of claims 82-122, wherein thereaction results in a yield of greater than about 50%.
 124. The methodof claim 123, wherein the yield is greater than about 75%.
 125. Themethod of either claim 123 or claim 124, wherein the yield is greaterthan about 90%.
 126. The method according to any one of claims 82-125,wherein the reaction produces a diastereomeric ratio of greater thanabout 1:1.
 127. The method of claim 126, wherein the diastereomericratio is greater than about 1.75:1.
 128. The method of either claim 126or claim 127, wherein the diastereomeric ratio is about 2:1.
 129. Themethod according to any one of claims 82-128, wherein the method furthercomprises forming a compound of the formula:

wherein: Y₄, Y₅, and Y₆ are each independently hydrogen, alkyl_((C≦8))or substituted alky_((C≦8)); R₁₆ and R₁₆′ are each independentlyhydrogen, alkoxy_((C≦8)), aralkoxy_((C≦12)), substituted alkoxy_((C≦8)),or substituted aralkoxy_((C≦12)); or R₁₆ and R₁₆′ are taken together andare alkoxydiyl_((C≦12)); R₁₇ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), or a substituted version ofany of these groups; or —C(O)OCH₂CH₂Si(CH₃)₃; X₃ is O, NH, S, or CH₂; X₄is O, NH, or S; X₅ is O, NH, or S; R₁₈ is hydrogen, alkyl_((C≦8)),alkenyl_((C≦8)), aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or asubstituted version of any of these groups; or —X₂—R₁₁, wherein: X₂ isalkanediyl_((C≦12)) or substituted alkanediyl_((C≦12)); and R₁₁ ishydroxy, amino, azido, carboxy, or cyano, alkenyl_((C≦6)),alkynyl_((C≦6)), heterocycloalkyl_((C≦12)), alkylamino_((C≦8)),dialkylamino_((C≦8)), alkoxy_((C≦8)), or a substituted version of any ofthese groups; R₁₉ is hydrogen, amino, carboxy, cyano, halo, hydroxy,nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; and R₂₀ and R₂₁ are eachindependently hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦8)), or a substituted version of any of thesegroups; or a salt, tautomer, or optical isomer thereof, comprisingreacting a compound of the formula:

wherein the variables are as defined above; with a metal catalyst and anoxidizing agent.
 130. The method of claim 129, wherein R₁₇ is nothydrogen, R₁₈ is not benzyl, R₁₉ is not hydrogen, and R₂₀ is not methyl.131. The method of either claim 129 or claim 130, wherein the metalcatalyst is a nickel(II) salt.
 132. The method of claim 131, wherein thenickel(II) salt is Ni(acac)₂.
 133. The method according to any one ofclaims 129-132, wherein the reaction comprises adding from about 0.01 toabout 1.0 equivalents of the metal catalyst relative to the compound offormula X.
 134. The method of claim 133, wherein the reaction comprisesadding from about 0.1 to about 0.5 equivalents of the metal catalyst.135. The method of either claim 133 or claim 134, wherein the reactioncomprises adding about 0.2 equivalents of the metal catalyst.
 136. Themethod according to any one of claims 129-135, wherein the oxidizingagent is a dioxirane compound.
 137. The method of claim 136, wherein theoxidizing agent is dimethyldioxirane (DMDO).
 138. The method accordingto any one of claims 129-137, wherein the reaction comprises adding fromabout 1.5 equivalents to about 10.0 equivalents of dimethyldioxiranerelative to the compound of formula X.
 139. The method of claim 138,wherein the reaction comprises adding from about 4.0 equivalents toabout 6.0 equivalents of dimethyldioxirane.
 140. The method of eitherclaim 138 or claim 139, wherein the reaction comprises adding about 5.1equivalents of dimethyldioxirane.
 141. The method according to any oneof claims 129-140, further comprising adding additionaldimethyldioxirane every two hours during the reaction.
 142. The methodof claim 141, wherein the additional dimethyldioxirane is about 1.5equivalents relative to the compound of formula X.
 143. The methodaccording to any one of claims 129-142, wherein the method furthercomprises a solvent.
 144. The method of claim 143, wherein the solventis an organic solvent.
 145. The method of either claim 143 or claim 144,wherein the solvent is a haloalkane_((C≦12)).
 146. The method of claim145, wherein the solvent is dichloromethane.
 147. The method accordingto any one of claims 129-146, wherein the reaction comprises running thereaction at a temperature from about −90° C. to about −40° C.
 148. Themethod of claim 147, wherein the temperature is from about −80° C. toabout −60° C.
 149. The method of either claim 147 or 148, wherein thetemperature is about −78° C.
 150. The method according to any one ofclaims 129-149, wherein the method further comprises allowing thereaction to warm to a temperature from about −80° C. to about −30° C.151. The method of claim 150, wherein the temperature is about −60° C.152. The method according to any one of claims 129-151, wherein thereaction is run for a time period from about 3 hours to about 12 hours.153. The method of claim 152, wherein the time period is from about 5hours to about 8 hours.
 154. The method of either claim 152 or claim153, wherein the time period is about 6.5 hours.
 155. The methodaccording to any one of claims 129-154, wherein the starting material isrecovered after the time period and subject to the reaction conditionsagain.
 156. The method according to any one of claims 129-155, whereinthe reaction results in a yield of greater than about 35% based uponrecovered starting material.
 157. The method of claim 156, wherein theyield is greater than about 50%.
 158. The method of either claim 156 orclaim 157, wherein the yield is greater than about 60%.
 159. The methodaccording to any one of claims 129-158, wherein the reaction produces adiastereomeric ratio of greater than about 1:1.
 160. The method of claim159, wherein the diastereomeric ratio is greater than about 1.75:1. 161.The method of either claim 159 or claim 160, wherein the diastereomericratio is about 2:1.
 162. The method according to any one of claims129-158, wherein the reaction further comprises reacting a compound offormula IX with a reducing agent to form a compound of the formula:

wherein: Y₄, Y₅, and Y₆ are each independently hydrogen, alkyl_((C≦8))or substituted alky_((C≦8)); R₁₆ and R₁₆′ are each independentlyhydrogen, alkoxy_((C≦8)), aralkoxy_((C≦12)), substituted alkoxy_((C≦8)),or substituted aralkoxy_((C≦12)); or R₁₆ and R₁₆′ are taken together andare alkoxydiyl_((C≦12)); R₁₇ is hydrogen, amino, carboxy, cyano, halo,hydroxy, nitro, or sulfo; alkyl_((C≦12)), alkoxy_((C≦12)),alkylamino_((C≦12)), dialkylamino_((C≦18)), or a substituted version ofany of these groups; or —C(O)OCH₂CH₂Si(CH₃)₃; X₃ is O, NH, S, or CH₂; X₄is O, NH, or S; R₁₈ is hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)),aryl_((C≦12)), aralkyl_((C≦12)), acyl_((C≦8)), or a substituted versionof any of these groups; or —X₂—R₁₁, wherein: X₂ is alkanediyl_((C≦12))or substituted alkanediyl_((C≦12)); and R₁₁ is hydroxy, amino, azido,carboxy, or cyano, alkenyl_((C≦6)), alkynyl_((C≦6)),heterocycloalkyl_((C≦12)), alkylamino_((C≦8)), dialkylamino_((C≦8)),alkoxy_((C≦8)), or a substituted version of any of these groups; R₁₉ ishydrogen, amino, carboxy, cyano, halo, hydroxy, nitro, or sulfo;alkyl_((C≦12)), alkoxy_((C≦12)), alkylamino_((C≦12)),dialkylamino_((C≦18)), amido_((C≦12)), or a substituted version of anyof these groups; or —NR₁₂C(O)R₁₃—NR₁₄R₁₅; wherein: R₁₂ is hydrogen,alkyl_((C≦8)), or substituted alkyl_((C≦8)); R₁₃ is alkanediyl_((C≦8))or substituted alkanediyl_((C≦8)); and R₁₄ and R₁₅ are eachindependently selected from: hydrogen, alkyl_((C≦12)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦12)), or a substituted version of any ofthese groups; or R₁₄ and R₁₅ are taken together and arealkanediyl_((C≦8)), alkoxydiyl_((C≦8)), alkylaminodiyl_((C≦8)), or asubstituted version of any of these groups; R₂₀ and R₂₁ are eachindependently hydrogen, alkyl_((C≦8)), alkenyl_((C≦8)), aryl_((C≦12)),aralkyl_((C≦12)), acyl_((C≦8)), or a substituted version of any of thesegroups; and R₂₃ is hydroxy, alkoxy_((C≦12)), or substitutedalkoxy_((C≦12)); or a salt, tautomer, or optical isomer thereof. 163.The method of claim 162, wherein the reducing agent is a soft hydridedonor.
 164. The method of claim 163, wherein the reducing agent is ametal borohydride reagent.
 165. The method of either claim 163 or claim164, wherein the reducing agent is sodium cyanoborohydride.
 166. Themethod according to any one of claims 162-165, wherein the reactioncomprises adding from about 5.0 equivalents to about 25.0 equivalents ofreducing agent relative to the compound of formula IX.
 167. The methodof claim 166, wherein the reaction comprises adding from about 8.0equivalents to about 12.0 equivalents of reducing agent.
 168. The methodof either claim 166 or claim 167, wherein the reaction comprises addingabout 10.0 equivalents of reducing agent.
 169. The method according toany one of claims 162-168, wherein the method further comprises asolvent.
 170. The method of claim 169, wherein the solvent is an organicsolvent.
 171. The method of either claim 169 or claim 170, wherein thesolvent is a ether_((C≦12)).
 172. The method of claim 171, wherein thesolvent is tetrahydrofuran.
 173. The method according to any one ofclaims 162-172, wherein the reaction comprises running the reaction at atemperature from about −90° C. to about −40° C.
 174. The method of claim173, wherein the temperature is from about −80° C. to about −60° C. 175.The method of either claim 173 or 174, wherein the temperature is about−78° C.
 176. The method according to any one of claims 162-175, whereinthe method further comprises allowing the reaction to warm to atemperature from about −80° C. to about −30° C.
 177. The method of claim176, wherein the temperature is about −60° C.
 178. The method accordingto any one of claims 162-177, wherein the reaction is run for a timeperiod from about 30 minutes to about 6 hours.
 179. The method of claim178, wherein the time period is from about 1 hour to about 3 hours. 180.The method of either claim 178 or claim 179, wherein the time period isabout 1.5 hours.
 181. The method according to any one of claims 162-180,wherein the reaction results in a yield of greater than about 20%. 182.The method of claim 181, wherein the yield is greater than about 25%.183. The method of either claim 181 or claim 182, wherein the yield isgreater than about 35%.
 184. The method according to any one of claims162-183, wherein the reaction produces a diastereomeric ratio of greaterthan about 1:1.
 185. The method of claim 184, wherein the diastereomericratio is greater than about 1.75:1.
 186. The method of either claim 184or claim 185, wherein the diastereomeric ratio is about 2:1.
 187. Themethod according to any one of claims 82-186, wherein the reactionfurther comprises purifying the reaction in a purification step. 188.The method of claim 187, wherein the purification method ischromatography.
 189. The method of either claim 187 or claim 188,wherein the purification method is column chromatography or highperformance liquid chromatography.